TREM COMPOSITIONS AND USES THEREOF

Abstract

The invention relates generally to tRNA-based effector molecules and methods relating thereto.

Claims

1. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, comprising: providing a mammalian host cell comprising an exogenous nucleic acid, e.g., a DNA or RNA, encoding the TREM; maintaining the mammalian cell under conditions sufficient to express the TREM; purifying the TREM from the mammalian host cell, e.g., according to a method described herein; and formulating the purified TREM as a pharmaceutical composition, e.g., by combining the TREM with a pharmaceutical excipient, thereby making the TREM pharmaceutical composition.

2. The method of claim 1, wherein the nucleic acid comprises an RNA, which upon reverse transcription, results in a DNA which can be transcribed into the TREM.

3. The method of claim 1 or 2, wherein the nucleic acid comprises an RNA sequence at least 90% identical to an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof.

4. The method of claim 1 or 2, wherein the nucleic acid comprises an RNA sequence comprising a consensus sequence provided herein.

5. The method of any one of the preceding claims, wherein the mammalian host cell is chosen from: a non-human cell or cell line, or a human cell or cell line, e.g., a HEK293T cell (e.g., a Freestyle 293-F cell), a HT-1080 cell, a PER.C6 cell, a HKB-11 cell, a CAP cell, a HuH-7 cell, a BHK 21 cell, an MRC-S cell, a MDCK cell, a VERO cell, a WI-38 cell, a Chinese Hamster Ovary (CHO) cell, or a MCF7 cell.

6. The method of any one of the preceding claims, wherein the purification step comprises one, two or all of the following steps, e.g., in the order recited: (i) separating nucleic acids from cellular debris to provide an RNA preparation; (ii) separating RNA of less than a threshold number of nucleotides, e.g., less than 500 nt, less than 400 nt, less than 300 nt, less than 250 nt, less than 200 nt, less than 150 nt, from larger RNA species in the RNA preparation to produce a small RNA preparation; and/or (iii) separating a TREM from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity-based separation.

7. A composition comprising a purified tRNA effector molecule (TREM) (e.g., a purified TREM composition made according to a method described herein), comprising: (i) an RNA sequence at least 90% identical to an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof; or (ii) an RNA sequence comprising a consensus sequence provided herein.

8. A GMP-grade, recombinant TREM composition (e.g., a TREM composition made in compliance with cGMP, and/or in accordance with similar requirements) comprising: (i) an RNA sequence at least 90% identical to an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof; or (ii) an RNA sequence comprising a consensus sequence provided herein.

9. The TREM composition of claim 7 or 8, wherein the composition comprises one or more, e.g., a plurality, of TREMs.

10. The TREM composition of any one of claims 7 to 9, wherein the composition comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 species of TREMs.

11. The TREM composition of any one of claims 7 to 10, wherein the TREM composition (or an intermediate in the production of a TREM composition) comprises one or more of the following characteristics: (i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%; (ii) host cell protein (HCP) contamination of less than 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, or 100 ng/ml; (iii) host cell protein (HCP) contamination of less than 0.1 ng, 1 ng, 5 ng, 10 ng, 15 ng, 20 ng, 25 ng, 30 ng, 35 ng, 40 ng, 50 ng, 60 ng, 70 ng, 80 ng, 90 ng, or 100 ng, per milligram (mg) of the TREM composition; (iv) DNA, e.g., host cell DNA, of less than 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, or 100 ng/ml; (v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% TREM fragments relative to full length TREMs; (vi) low levels or absence of endotoxins, e.g., a negative result as measured by the Limulus amebocyte lysate (LAL) test; (vii) in-vitro translation activity, e.g., as measured by an assay described in Example 15; (viii) TREM concentration of at least 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 0.1 ug/mL, 0.5 ug/mL, 1 ug/mL, 2 ug/mL, 5 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, 40 ug/mL, 50 ug/mL, 60 ug/mL, 70 ug/mL, 80 ug/mL, 100 ug/mL, 200 ug/mL, 300 ug/mL, 500 ug/mL, 1000 ug/mL, 5000 ug/mL, 10,000 ug/mL, or 100,000 ug/mL; (ix) sterility, e.g., as per cGMP guidelines for sterile drug products, e.g., the composition or preparation supports the growth of fewer than 100 viable microorganisms as tested under aseptic conditions, the composition or preparation meets the standard of USP <71>, and/or the composition or preparation meets the standard of USP <85>; or (x) viral contamination, e.g., the composition or preparation has an absence of, or an undetectable level of viral contamination.

12. A method of modulating a tRNA pool in a cell comprising: providing a purified TREM composition, and contacting the cell with the TREM composition, thereby modulating the tRNA pool in the cell.

13. The method of claim 12, wherein the TREM composition is made by: providing a mammalian host cell comprising an exogenous nucleic acid, e.g., a DNA or RNA, encoding the TREM; maintaining the mammalian cell under conditions sufficient to express the TREM; and/or purifying the TREM from the mammalian host cell, e.g., according to a method described herein.

14. The method of claim 12 or 13, wherein the mammalian host cell is chosen from: a non-human cell or cell line, or a human cell or cell line, e.g., a HEK293T cell (e.g., a Freestyle 293-F cell), a HT-1080 cell, a PER.C6 cell, a HKB-11 cell, a CAP cell, a HuH-7 cell, a BHK 21 cell, an MRC-S cell, a MDCK cell, a VERO cell, a WI-38 cell, a Chinese Hamster Ovary (CHO) cell, or a MCF7 cell.

15. The method of any one of claims 12 to 14, wherein the purification step comprises one, two or all of the following steps, e.g., in the order recited: (i) separating nucleic acids from cellular debris to provide an RNA preparation; (ii) separating RNA of less than a threshold number of nucleotides, e.g., less than 500 nt, less than 400 nt, less than 300 nt, less than 250 nt, less than 200 nt, less than 150 nt, from larger RNA species in the RNA preparation to produce a small RNA preparation; and/or (iii) separating a TREM from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity-based separation.

16. The method of any one of claims 12 to 15, wherein the TREM comprises: (i) an RNA sequence at least 80% identical to an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof; or (ii) an RNA sequence comprising a consensus sequence provided herein.

17. A method of making a tRNA effector molecule (TREM) composition, comprising: (a) providing a mammalian host cell comprising exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM under conditions sufficient to express the TREM, and (b) purifying the expressed TREM from the mammalian host cell to produce a TREM composition, thereby making the TREM composition.

18. A method of making a pharmaceutical TREM composition comprising: combining a) a TREM, e.g., a purified TREM composition, e.g., a TREM composition made by a method described herein; and b) a pharmaceutically acceptable component, e.g., an excipient, thereby making a pharmaceutical TREM composition.

19. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, comprising: purifying the TREM from a mammalian host cell; formulating the purified TREM as a pharmaceutical composition, e.g., by combining the TREM with a pharmaceutical excipient, thereby making the TREM pharmaceutical composition.

20. A method of making a TREM composition, comprising: contacting a TREM containing a reaction mixture with a reagent, e.g., a capture reagent or a separation reagent, comprising a nucleic acid sequence complimentary with a TREM; thereby making a TREM composition.

21. A method of making a pharmaceutical composition, comprising: a) providing a purified TREM composition, e.g., a purified TREM composition made by culturing a mammalian host cell comprising DNA or RNA encoding a TREM under conditions sufficient to express the TREM, and purifying the expressed TREM from the host cell culture to produce a purified TREM composition, b) providing a value, e.g., by evaluating or testing, for one or more of the following characteristics of the purified TREM composition: (i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%; (ii) host cell protein (HCP) contamination of less than 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, or 100 ng/ml; (iii) host cell protein (HCP) contamination of less than 0.1 ng, 1 ng, 5 ng, 10 ng, 15 ng, 20 ng, 25 ng, 30 ng, 35 ng, 40 ng, 50 ng, 60 ng, 70 ng, 80 ng, 90 ng, or 100 ng per milligram (mg) of the TREM composition; (iv) DNA, e.g., host cell DNA, of less than 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, or 100 ng/ml; (v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% TREM fragments relative to full length TREMs; (vi) low levels or absence of endotoxins, e.g., a negative result as measured by the Limulus amebocyte lysate (LAL) test; (vii) in-vitro translation activity, e.g., as measured by an assay described in Example 15; (viii) TREM concentration of at least 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 0.1 ug/mL, 0.5 ug/mL, 1 ug/mL, 2 ug/mL, 5 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, 40 ug/mL, 50 ug/mL, 60 ug/mL, 70 ug/mL, 80 ug/mL, 100 ug/mL, 200 ug/mL, 300 ug/mL, 500 ug/mL, 1000 ug/mL, 5000 ug/mL, 10,000 ug/mL, or 100,000 ug/mL; (ix) sterility, e.g., as per cGMP guidelines for sterile drug products, e.g., the composition or preparation supports the growth of fewer than 100 viable microorganisms as tested under aseptic conditions, the composition or preparation meets the standard of USP <71>, and/or the composition or preparation meets the standard of USP <85>; or (x) viral contamination, e.g., the composition or preparation has an absence of, or an undetectable level of viral contamination. c) optionally, formulating the purified TREM composition as a pharmaceutical drug product (e.g., combining the TREM composition with a pharmaceutical excipient) if it meets a reference criteria for the one or more characteristics, thereby making a pharmaceutical composition.

22. A pharmaceutical tRNA effector molecule (TREM) composition, comprising (i) an RNA sequence at least 80% identical to an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof; or (ii) an RNA sequence comprising a consensus sequence provided herein.

23. A recombinant TREM composition of at least 0.5 g, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, 10 g, 15 g, 20 g, 30 g, 40 g, 50 g, 100 g, 200 g, 300 g, 400 g or 500 g.

24. A recombinant TREM composition of between 0.5 g to 500 g, between 0.5 g to 400 g, between 0.5 g to 300 g, between 0.5 g to 200 g, between 0.5 g to 100 g, between 0.5 g to 50 g, between 0.5 g to 40 g, between 0.5 g to 30 g, between 0.5 g to 20 g, between 0.5 g to 10 g, between 0.5 g to 9 g, between 0.5 g to 8 g, between 0.5 g to 7 g, between 0.5 g to 6 g, between 0.5 g to 5 g, between 0.5 g to 4 g, between 0.5 g to 3 g, between 0.5 g to 2 g, between 0.5 g to 1 g, between 1 g to 500 g, between 2 g to 500 g, between 5 g to 500 g, between 10 g to 500 g, between 20 g to 500 g, between 30 g to 500 g, between 40 g to 500 g, between 50 g to 500 g, between 100 g to 500 g, between 200 g to 500 g, between 300 g to 500 g, or between 400 g to 500 g.

25. A TREM composition comprising a consensus sequence of Formula I.sub.ZZZ, R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72 wherein: R is a ribonucleotide residue; (i) .sub.ZZZ indicates any of the twenty amino acids; (ii) Formula I corresponds to all species; and (iii) x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1- 24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271).

26. A TREM composition comprising a consensus sequence of Formula II.sub.ZZZ, R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72 wherein: R is a ribonucleotide residue; (i) .sub.ZZZ indicates any of the twenty amino acids; (ii) Formula II corresponds to mammals; and (iii) x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1- 24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271).

27. A TREM composition comprising a consensus sequence of Formula III.sub.ZZZ, R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72 wherein: R is a ribonucleotide residue; (i) .sub.ZZZ indicates any of the twenty amino acids; (ii) Formula III corresponds to humans; and (iii) x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1- 24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271).

28. A method of contacting a cell, tissue, or subject with a TREM, comprising contacting the cell, tissue or subject with a purified TREM composition, thereby contacting a cell, tissue, or subject with the TREM.

29. A method of presenting a TREM to a cell, tissue, or subject with a TREM, comprising contacting the cell, tissue or subject with a purified TREM composition, thereby presenting the TREM to a cell, tissue, or subject.

30. A method of forming a TREM-contacted cell, tissue, or subject, comprising contacting the cell, tissue or subject with a purified TREM composition, thereby forming a TREM-contacted cell, tissue, or subject.

31. A method of using a TREM comprising, contacting the cell, tissue or subject with a purified TREM composition, thereby using the TREM.

32. A method of applying a TREM to a cell, tissue, or subject, comprising contacting the cell, tissue or subject with a purified TREM composition, thereby applying a TREM to a cell, tissue, or subject.

33. A method of exposing a cell, tissue, or subject to a TREM, comprising contacting the cell, tissue or subject with a purified TREM composition, thereby exposing a cell, tissue, or subject to a TREM.

34. A method of forming an admixture of a TREM and a cell, tissue, or subject, comprising contacting the cell, tissue or subject with a TREM composition, thereby forming an admixture of a TREM and a cell, tissue, or subject.

35. A method of delivering a TREM to a cell, tissue, or subject, comprising: providing a cell, tissue, or subject, and contacting the cell, tissue, or subject, with a TREM composition, e.g., a purified TREM composition, e.g., a pharmaceutical TREM composition.

36. A method, e.g., an ex vivo method, of modulating the metabolism, e.g., the translational capacity of an organelle, comprising: providing a preparation of an organelle, e.g., mitochondria or chloroplasts, and contacting the organelle with a pharmaceutical TREM composition.

37. A method of treating a subject, e.g., modulating the metabolism, e.g., the translational capacity of a cell, in a subject, comprising: providing, e.g., administering to the subject, an exogenous nucleic acid, e.g., a DNA or RNA, which encodes a TREM, thereby treating the subject.

38. A cell comprising a TREM made according to a method of making a TREM disclosed herein.

39. A cell comprising a TREM disclosed herein.

40. A cell comprising an exogenous nucleic acid comprising: a nucleic acid sequence, e.g., DNA or RNA, that encodes a TREM, wherein the nucleic acid sequence comprises: (i) a control region sequence; (ii) a sequence encoding a modified TREM; (iii) a sequence encoding more than one TREM; (iv) a sequence other than a tRNA.sup.Met sequence; or (v) a promoter sequence that comprises a Pol III recognition site, e.g., a U6 promoter, a 7SK promoter or a H1 promoter, or a fragment thereof.

41. A reaction mixture comprising a TREM and a reagent, e.g., a capture reagent, or a separation reagent.

42. A bioreactor comprising a plurality of mammalian host cells described herein comprising exogenous DNA or RNA.

43. A master cell bank comprising a host cell, e.g., as described herein.

44. A method of evaluating a composition of TREM, e.g., a GMP-grade TREM (i.e., a TREM made in compliance with cGMP, and/or in accordance with similar requirements), comprising acquiring a value for one or more of the following characteristics of the purified TREM composition: (i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%; (ii) host cell protein (HCP) contamination of less than 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, or 100 ng/ml; (iii) host cell protein (HCP) contamination of less than 0.1 ng, 1 ng, 5 ng, 10 ng, 15 ng, 20 ng, 25 ng, 30 ng, 35 ng, 40 ng, 50 ng, 60 ng, 70 ng, 80 ng, 90 ng, or 100 ng per milligram (mg) of the TREM composition; (iv) DNA, e.g., host cell DNA, of less than 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, or 100 ng/ml; (v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% TREM fragments relative to full length TREMs; (vi) low levels or absence of endotoxins, e.g., a negative result as measured by the Limulus amebocyte lysate (LAL) test; (vii) in-vitro translation activity, e.g., as measured by an assay described in Example 15; (viii) TREM concentration of at least 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 0.1 ug/mL, 0.5 ug/mL, 1 ug/mL, 2 ug/mL, 5 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, 40 ug/mL, 50 ug/mL, 60 ug/mL, 70 ug/mL, 80 ug/mL, 100 ug/mL, 200 ug/mL, 300 ug/mL, 500 ug/mL, 1000 ug/mL, 5000 ug/mL, 10,000 ug/mL, or 100,000 ug/mL; (ix) sterility, e.g., the composition or preparation supports the growth of fewer than 100 viable microorganisms as tested under aseptic conditions, the composition or preparation meets the standard of USP <71>, and/or the composition or preparation meets the standard of USP <85> as described by cGMP guidelines for sterile drug products produced by aseptic processing; or (x) viral contamination, e.g., the composition or preparation has an absence of, or an undetectable level of viral contamination.

Description

BRIEF DESCRIPTIONS OF THE DRAWINGS

[0319] FIGS. 1A-1C are graphs showing an increase in cell growth in three cells lines after transfection with a TREM corresponding to the initiator methionine (iMet). FIG. 1A is a graph showing increased % cellular confluency (a measure of cell growth) of U20S cells transfected with Cy3-labeled iMet-CAT-TREM or transfected with a Cy3-labeled non-targeted control. FIG. 1B is a graph showing increased % cellular confluency (a measure of cell growth) of H1299 cells transfected with Cy3-labeled iMet-CAT-TREM or transfected with a Cy3-labeled non-targeted control. FIG. 1C is a graph showing increased % cellular confluency (a measure of cell growth) of Hela cells transfected with Cy3-labeled iMet-CAT-TREM or transfected with a Cy3-labeled non-targeted control.

[0320] FIG. 2 is a graph depicting an increase in NanoLuc reporter expression upon addition of iMET-TREM to a translational reaction with cell free lysate. As a control, a translational reaction with buffer was performed.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

[0321] The present disclosure features tRNA-based effector molecules (TREMs) and methods relating thereto. As disclosed herein tRNA-based effector molecules (TREMs) are complex molecules which can mediate a variety of cellular processes. Pharmaceutical TREM compositions can be administered to a cell, a tissue, or to a subject to modulate these functions.

Definitions

[0322] A “cognate adaptor function TREM,” as that term is used herein, refers to a TREM which mediates initiation or elongation with the AA (the cognate AA) associated in nature with the anti-codon of the TREM.

[0323] “Decreased expression,” as that term is used herein, refers to a decrease in comparison to a reference, e.g., in the case where altered control region, or addition of an agent, results in a decreased expression of the subject product, it is decreased relative to an otherwise similar cell without the alteration or addition.

[0324] An “exogenous nucleic acid,” as that term is used herein, refers to a nucleic acid sequence that is not present in or differs by at least one nucleotide from the closest sequence in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced. In an embodiment, an exogenous nucleic acid comprises a nucleic acid that encodes a TREM.

[0325] An “exogenous TREM,” as that term is used herein, refers to a TREM that:

[0326] (a) differs by at least one nucleotide or one post transcriptional modification from the closest sequence tRNA in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced;

[0327] (b) has been introduced into a cell other than the cell in which it was transcribed;

[0328] (c) is present in a cell other than one in which it naturally occurs; or

[0329] (d) has an expression profile, e.g., level or distribution, that is non-wildtype, e.g., it is expressed at a higher level than wildtype. In an embodiment, the expression profile can be mediated by a change introduced into a nucleic acid that modulates expression or by addition of an agent that modulates expression of the RNA molecule. In an embodiment an exogenous TREM comprises 1, 2, 3 or 4 of properties (a)-(d).

[0330] A “GMP-grade composition,” as that term is used herein, refers to a composition in compliance with current good manufacturing practice (cGMP) guidelines, or other similar requirements. In an embodiment, a GMP-grade composition can be used as a pharmaceutical product.

[0331] As used herein, the terms “increasing” and “decreasing” refer to modulating that results in, respectively, greater or lesser amounts of function, expression, or activity of a particular metric relative to a reference. For example, subsequent to administration to a cell, tissue or subject of a TREM described herein, the amount of a marker of a metric (e.g., protein translation, mRNA stability, protein folding) as described herein may be increased or decreased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98%, 2×, 3×, 5×, 10× or more relative to the amount of the marker prior to administration or relative to the effect of a negative control agent. The metric may be measured subsequent to administration at a time that the administration has had the recited effect, e.g., at least 12 hours, 24 hours, one week, one month, 3 months, or 6 months, after a treatment has begun.

[0332] “Increased expression,” as that term is used herein, refers to an increase in comparison to a reference, e.g., in the case where altered control region, or addition of an agent, results in an increased expression of the subject product, it is increased relative to an otherwise similar cell without the alteration or addition.

[0333] A “non-cognate adaptor function TREM,” as that term is used herein, refers to a TREM which mediates initiation or elongation with an AA (a non-cognate AA) other than the AA associated in nature with the anti-codon of the TREM. In an embodiment, a non-cognate adaptor function TREM is also referred to as a mischarged TREM (mTREM).

[0334] A “non-naturally occurring sequence,” as that term is used herein, refers to a sequence wherein an Adenine is replaced by a residue other than an analog of Adenine, a Cytosine is replaced by a residue other than an analog of Cytosine, a Guanine is replaced by a residue other than an analog of Guanine, and a Uracil is replaced by a residue other than an analog of Uracil. An analog refers to any possible derivative of the ribonucleotides, A, G, C or U. In an embodiment, a sequence having a derivative of any one of ribonucleotides A, G, C or U is a non-naturally occurring sequence.

[0335] An “oncogene,” as that term is used herein, refers to a gene that modulates one or more cellular processes including: cell fate determination, cell survival and genome maintenance. In an embodiment, an oncogene provides a selective growth advantage to the cell in which it is present, e.g., deregulated, e.g., genetically deregulated (e.g., mutated or amplified) or epigenetically deregulated. Exemplary oncogenes include, Myc (e.g., c-Myc, N-Myc or L-Myc), c-Jun, Wnt, or RAS.

[0336] A “pharmaceutical TREM composition,” as that term is used herein, refers to a TREM composition that is suitable for pharmaceutical use. Typically, a pharmaceutical TREM composition comprises a pharmaceutical excipient. In an embodiment the TREM will be the only active ingredient in the pharmaceutical TREM composition. In embodiments the pharmaceutical TREM composition is free, substantially free, or has less than a pharmaceutically acceptable amount, of host cell proteins, DNA, e.g., host cell DNA, endotoxins, and bacteria.

[0337] A “post-transcriptional processing,” as that term is used herein, with respect to a subject molecule, e.g., a TREM, RNA or tRNAs, refers to a covalent modification of the subject molecule. In an embodiment, the covalent modification occurs post-transcriptionally. In an embodiment, the covalent modification occurs co-transcriptionally. In an embodiment the modification is made in vivo, e.g., in a cell used to produce a TREM. In an embodiment the modification is made ex vivo, e.g., it is made on a TREM isolated or obtained from the cell which produced the TREM. In an embodiment, the post-transcriptional modification is selected from a post-transcriptional modification listed in Table 2.

[0338] A “recombinant TREM,” as that term is used herein, refers to a TREM that was expressed in a cell modified by human intervention, having a modification that mediates the production of the TREM, e.g., the cell comprises an exogenous sequence encoding the TREM, or a modification that mediates expression, e.g., transcriptional expression or post-transcriptional modification, of the TREM. A recombinant TREM can have the same, or a different, sequence, set of post-transcriptional modifications, or tertiary structure, as a reference tRNA, e.g., a native tRNA.

[0339] A “synthetic TREM,” as that term is used herein, refers to a TREM which was synthesized other than in a cell having an endogenous nucleic acid encoding the TREM, e.g., by cell-free solid phase synthesis. A synthetic TREM can have the same, or a different, sequence, set of post-transcriptional modifications, or tertiary structure, as a native tRNA.

[0340] A “TREM expressed in a heterologous cell,” as that term is used herein, refers to a TREM made under non-native conditions. E.g., a TREM, i) made in a cell that, differs, e.g., genetically, metabolically (e.g., has a different profile of gene expression or has a different level of a cellular component, e.g., an absorbed nutrient), or epigenetically, from a naturally occurring cell; ii) made in a cell that, is cultured under conditions, e.g., nutrition, pH, temperature, cell density, or stress conditions, that are different from native conditions (native conditions are the conditions under which a cell makes a tRNA in nature); or iii) was made in a cell at a level, at a rate, or at a concentration, or was localized in a compartment or location, that differs from a reference, e.g., at a level, at a rate, or at a concentration, or was localized in a compartment or location, that differs from that which occurs under native conditions. A TREM expressed in a heterologous cell can have the same, or a different, sequence, set of post-transcriptional modifications, or tertiary structure, as a native tRNA.

[0341] A “tRNA”, as that term is used herein, refers to a naturally occurring transfer ribonucleic acid in its native state.

[0342] A “tRNA-based effector molecule” or “TREM,” as that term is used herein, refers to an RNA molecule comprising a structure or property from (a)-(v) below, and which is a recombinant TREM, a synthetic TREM, or a TREM expressed from a heterologous cell. A TREM can have a plurality (e.g., 2, 3, 4, 5, 6, 7, 8, 9) of the structures and functions of (a)-(v).

[0343] In an embodiment, a TREM is non-native, as evaluated by structure or the way in which it was made.

[0344] In an embodiment, a TREM comprises one or more of the following structures or properties:

[0345] (a′) an optional linker region of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 1 region;

[0346] (a) an amino acid attachment domain that binds an amino acid, e.g., an acceptor stem domain (AStD), wherein an AStD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, acceptance of an amino acid, e.g., its cognate amino acid or a non-cognate amino acid, and transfer of the amino acid (AA) in the initiation or elongation of a polypeptide chain. Typically, the AStD comprises a 3′-end adenosine (CCA) for acceptor stem charging which is part of synthetase recognition. In an embodiment the AStD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring AStD, e.g., an AStD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of an AStD, e.g., an AStD encoded by a nucleic acid in Table 1, which fragment in embodiments has AStD activity and in other embodiments does not have AStD activity. (One of ordinary skill can determine the relevant corresponding sequence for any of the domains, stems, loops, or other sequence features mentioned herein from a sequence encoded by a nucleic acid in Table 1. E.g., one of ordinary skill can determine the sequence which corresponds to an AStD from a tRNA sequence encoded by a nucleic acid in Table 1.)

[0347] In an embodiment the AStD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

[0348] In an embodiment, the AStD comprises residues R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7 and residues R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71 of Formula I.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the AStD comprises residues R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7 and residues R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71 of Formula II.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the AStD comprises residues R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7 and residues R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71 of Formula III.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids;

[0349] (a′-1) a linker comprising residues R.sub.8-R.sub.9 of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 2 region;

[0350] (b) a dihydrouridine hairpin domain (DHD), wherein a DHD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, recognition of aminoacyl-tRNA synthetase, e.g., acts as a recognition site for aminoacyl-tRNA synthetase for amino acid charging of the TREM. In embodiments, a DHD mediates the stabilization of the TREM's tertiary structure. In an embodiment the DHD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring DHD, e.g., a DHD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a DHD, e.g., a DHD encoded by a nucleic acid in Table 1, which fragment in embodiments has DHD activity and in other embodiments does not have DHD activity.

[0351] In an embodiment the DHD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

[0352] In an embodiment, the DHD comprises residues R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14 R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28 of Formula I.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the DHD comprises residues R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14 R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28 of Formula II.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the DHD comprises residues R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14 R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28 of Formula III.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids;

[0353] (b′-1) a linker comprising residue R.sub.29 of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 3 region;

[0354] (c) an anticodon that binds a respective codon in an mRNA, e.g., an anticodon hairpin domain (ACHD), wherein an ACHD comprises sufficient sequence, e.g., an anticodon triplet, to mediate, e.g., when present in an otherwise wildtype tRNA, pairing (with or without wobble) with a codon; In an embodiment the ACHD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of an ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1, which fragment in embodiments has ACHD activity and in other embodiments does not have ACHD activity.

[0355] In an embodiment the ACHD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

[0356] In an embodiment, the ACHD comprises residues -R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46 of Formula I.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids;

[0357] In an embodiment, the ACHD comprises residues -R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46 of Formula II.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the ACHD comprises residues -R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46 of Formula III.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids;

[0358] (d) a variable loop domain (VLD), wherein a VLD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, recognition of aminoacyl-tRNA synthetase, e.g., acts as a recognition site for aminoacyl-tRNA synthetase for amino acid charging of the TREM. In embodiments, a VLD mediates the stabilization of the TREM's tertiary structure. In an embodiment, a VLD modulates, e.g., increases, the specificity of the TREM, e.g., for its cognate amino acid, e.g., the VLD modulates the TREM's cognate adaptor function. In an embodiment the VLD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring VLD, e.g., a VLD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a VLD, e.g., a VLD encoded by a nucleic acid in Table 1, which fragment in embodiments has VLD activity and in other embodiments does not have VLD activity.

[0359] In an embodiment the VLD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section.

[0360] In an embodiment, the VLD comprises residue -[R.sub.47].sub.x of a consensus sequence provided in the “Consensus Sequence” section, wherein x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271);

[0361] (e) a thymine hairpin domain (THD), wherein a THD comprises sufficient RNA sequence, to mediate, e.g., when present in an otherwise wildtype tRNA, recognition of the ribosome, e.g., acts as a recognition site for the ribosome to form a TREM-ribosome complex during translation. In an embodiment the THD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring THD, e.g., a THD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a THD, e.g., a THD encoded by a nucleic acid in Table 1, which fragment in embodiments has THD activity and in other embodiments does not have THD activity.

[0362] In an embodiment the THD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

[0363] In an embodiment, the THD comprises residues -R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64 of Formula I.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the THD comprises residues -R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64 of Formula II.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the THD comprises residues -R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64 of Formula III.sub.ZZZ, wherein ZZZ indicates any of the twenty amino acids;

[0364] (e′1) a linker comprising residue R.sub.72 of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 4 region;

[0365] (f) under physiological conditions, it comprises a stem structure and one or a plurality of loop structures, e.g., 1, 2, or 3 loops. A loop can comprise a domain described herein, e.g., a domain selected from (a)-(e). A loop can comprise one or a plurality of domains. In an embodiment, a stem or loop structure has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1, which fragment in embodiments has activity of a stem or loop structure, and in other embodiments does not have activity of a stem or loop structure;

[0366] (g) a tertiary structure, e.g., an L-shaped tertiary structure;

[0367] (h) adaptor function, i.e., the TREM mediates acceptance of an amino acid, e.g., its cognate amino acid and transfer of the AA in the initiation or elongation of a polypeptide chain;

[0368] (i) cognate adaptor function wherein the TREM mediates acceptance and incorporation of an amino acid (e.g., cognate amino acid) associated in nature with the anti-codon of the TREM to initiate or elongate a polypeptide chain;

[0369] (j) non-cognate adaptor function, wherein the TREM mediates acceptance and incorporation of an amino acid (e.g., non-cognate amino acid) other than the amino acid associated in nature with the anti-codon of the TREM in the initiation or elongation of a polypeptide chain;

[0370] (k) a regulatory function, e.g., an epigenetic function (e.g., gene silencing function or signaling pathway modulation function), cell fate modulation function, mRNA stability modulation function, protein stability modulation function, protein transduction modulation function, or protein compartmentalization function;

[0371] (l) a structure which allows for ribosome binding;

[0372] (m) a post-transcriptional modification, e.g., it comprises one or more modifications from Table 2, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 modifications listed in Table 2;

[0373] (n) the ability to inhibit a functional property of a tRNA, e.g., any of properties (h)-(k) possessed by a tRNA;

[0374] (o) the ability to modulate cell fate;

[0375] (p) the ability to modulate ribosome occupancy;

[0376] (q) the ability to modulate protein translation;

[0377] (r) the ability to modulate mRNA stability;

[0378] (s) the ability to modulate protein folding and structure;

[0379] (t) the ability to modulate protein transduction or compartmentalization;

[0380] (u) the ability to modulate protein stability; or

[0381] (v) the ability to modulate a signaling pathway, e.g., a cellular signaling pathway.

[0382] In an embodiment, a TREM comprises a full-length tRNA molecule or a fragment thereof.

[0383] In an embodiment, a TREM comprises the following properties: (a)-(e).

[0384] In an embodiment, a TREM comprises the following properties: (a) and (c).

[0385] In an embodiment, a TREM comprises the following properties: (a), (c) and (h).

[0386] In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (b).

[0387] In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (e).

[0388] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (b) and (e).

[0389] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (b), (e) and (g).

[0390] In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (m).

[0391] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), and (g).

[0392] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m) and (b).

[0393] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m) and (e).

[0394] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), (g), (b) and (e).

[0395] In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), (g), (b), (e) and (q).

[0396] In an embodiment, a TREM comprises:

[0397] (i) an amino acid attachment domain that binds an amino acid (e.g., an AStD, as described in (a) herein; and

[0398] (ii) an anticodon that binds a respective codon in an mRNA (e.g., an ACHD, as described in (c) herein).

[0399] In an embodiment the TREM comprises a flexible RNA linker which provides for covalent linkage of (i) to (ii).

[0400] In an embodiment, the TREM mediates protein translation.

[0401] In an embodiment a TREM comprises a linker, e.g., an RNA linker, e.g., a flexible RNA linker, which provides for covalent linkage between a first and a second structure or domain. In an embodiment, an RNA linker comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 ribonucleotides. A TREM can comprise one or a plurality of linkers, e.g., in embodiments a TREM comprising (a), (b), (c), (d) and (e) can have a first linker between a first and second domain, and a second linker between a third domain and another domain.

[0402] In an embodiment, a TREM comprises an RNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with, or which differs by no more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 ribonucleotides from, an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises a TREM domain, e.g., a domain described herein, comprising at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical with, or which differs by no more than 1, 2, 3, 4, 5, 10, or 15, ribonucleotides from, an RNA encoded by a DNA sequence listed in Table 1, or a fragment or a functional fragment thereof. In an embodiment, a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by DNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with a DNA sequence listed in Table 1, or a fragment or functional fragment thereof.

[0403] In an embodiment, a TREM is 76-90 nucleotides in length. In embodiments, a TREM or a fragment or functional fragment thereof is between 10-90 nucleotides, between 10-80 nucleotides, between 10-70 nucleotides, between 10-60 nucleotides, between 10-50 nucleotides, between 10-40 nucleotides, between 10-30 nucleotides, between 10-20 nucleotides, between 20-90 nucleotides, between 20-80 nucleotides, 20-70 nucleotides, between 20-60 nucleotides, between 20-50 nucleotides, between 20-40 nucleotides, between 30-90 nucleotides, between 30-80 nucleotides, between 30-70 nucleotides, between 30-60 nucleotides, or between 30-50 nucleotides.

[0404] In an embodiment, a TREM is aminoacylated, e.g., charged, with an amino acid by an aminoacyl tRNA synthetase.

[0405] In an embodiment, a TREM is not charged with an amino acid, e.g., an uncharged TREM (uTREM).

[0406] In an embodiment, a TREM comprises less than a full length tRNA. In embodiments, a TREM can correspond to a naturally occurring fragment of a tRNA, or to a non-naturally occurring fragment. Exemplary fragments include: TREM halves (e.g., from a cleavage in the ACHD, e.g., in the anticodon sequence, e.g., 5′halves or 3′ halves); a 5′ fragment (e.g., a fragment comprising the 5′ end, e.g., from a cleavage in a DHD or the ACHD); a 3′ fragment (e.g., a fragment comprising the 3′ end, e.g., from a cleavage in the THD); or an internal fragment (e.g., from a cleavage in one or more of the ACHD, DHD or THD).

[0407] A “TREM composition,” as that term is used herein, refers to a composition comprising a plurality of TREMs. A TREM composition can comprise one or more species of TREMs. In an embodiment, the composition comprises only a single species of TREM. In an embodiment, the TREM composition comprises a first TREM species and a second TREM species. In an embodiment, the TREM composition comprises X TREM species, wherein X=2, 3, 4, 5, 6, 7, 8, 9, or 10. In an embodiment, the TREM has at least 70, 75, 80, 85, 90, or 95, or has 100%, identity with a sequence encoded by a nucleic acid in Table 1. A TREM composition can comprise one or more species of TREMs. In an embodiment, the TREM composition is purified from cell culture. In an embodiment the cell culture from which the TREM is purified comprises at least 1×10.sup.7 host cells, 1×10.sup.8 host cells, 1×10.sup.9 host cells, 1×10.sup.10 host cells, 1×10.sup.11 host cells, 1×10.sup.12 host cells, 1×10.sup.13 host cells, or 1×10.sup.14 host cells. In an embodiment, the TREM composition is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% dry weight TREMs (for a liquid composition dry weight refers to the weight after removal of substantially all liquid, e.g., after lyophilization). In an embodiment, the composition is a liquid. In an embodiment, the composition is dry, e.g., a lyophilized material. In an embodiment, the composition is a frozen composition. In an embodiment, the composition is sterile. In an embodiment, the composition comprises at least 0.5 g, 1.0 g, 5.0 g, 10 g, 15 g, 25 g, 50 g, 100 g, 200 g, 400 g, or 500 g (e.g., as determined by dry weight) of TREM.

[0408] A “tumor suppressor,” as that term is used herein, refers to a gene that modulates one or more cellular processes including: cell fate determination, cell survival and genome maintenance. In an embodiment, a tumor suppressor provides a selective growth advantage to the cell in which it is deregulated, e.g., genetically deregulated (e.g., mutated or deleted) or epigenetically deregulated. Exemplary tumor suppressors include p53 or Rb.

Host Cells

[0409] A host cell is a cell (e.g., a cultured cell) that can be used for expression and/or purification of a TREM. In an embodiment, a host cell comprises a mammalian cell, e.g., a human cell. In an embodiment, a host cell comprises a non-mammalian cell, e.g., a yeast cell. In an embodiment, a host cell comprises a HeLa cell, a HEK293T cell (e.g., a Freestyle 293-F cell), a HT-1080 cell, a PER.C6 cell, a HKB-11 cell, a CAP cell, a HuH-7 cell, a BHK 21 cell, an MRC-S cell, a MDCK cell, a VERO cell, a WI-38 cell, or a Chinese Hamster Ovary (CHO) cell. In an embodiment, a host cell comprises a cancer cell, e.g., a solid tumor cell (e.g., a breast cancer cell (e.g., a MCF7 cell), a pancreatic cell line (e.g. a MIA PaCa-2 cell), a lung cancer cell, or a prostate cancer cell, or a hematological cancer cell). In an embodiment, a host cell comprises a cell that expresses one or more tissue-specific tRNAs. For example, a host cell can comprise a cell derived from a tissue associated with expression of a tRNA, e.g., a tissue-specific tRNA. In an embodiment, a host cell that expresses a tissue-specific tRNA is modified to express a TREM, or a fragment thereof.

[0410] In an embodiment, the host cell is not a bacterial cell, e.g., an E. coli cell.

[0411] In an embodiment, a host cell is a cell that can be maintained under conditions that allow for expression of a TREM.

[0412] In an embodiment, a host cell is capable of post-transcriptionally modifying the TREM, e.g., adding a post-transcriptional modification selected from Table 2. In an embodiment, a host cell expresses (e.g., naturally or heterologously) an enzyme listed in Table 2. In an embodiment, a host cell expresses (e.g., naturally or heterologously) an enzyme, e.g., an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., or one or more of Dicer, Angiogenin, RNaseA, RNaseP, RNaseZ, Rny1 or PrrC.

Method of Culturing Host Cell

[0413] A host cell can be cultured in a medium that promotes growth, e.g., proliferation or hyperproliferation of the host cell. A host cell can be cultured in a suitable media, e.g., any of the following media: DMEM, MEM, MEM alpha, RPMI, F-10 media, F-12 media, DMEM/F-12 media, IMDM, Medium 199, Leibovitz L-15, McCoys's 5A, MDCB media, or CMRL media. In an embodiment the media is supplemented with glutamine. In an embodiment, the media is not supplemented with glutamine. In an embodiment, a host cell is cultured in media that has an excess of nutrients, e.g., is not nutrient limiting. A host cell can be cultured in a medium comprising or supplemented with one or a combination of growth factors, cytokines or hormones, e.g., one or a combination of serum (e.g., fetal bovine serum (FBS)), HEPES, fibroblast growth factor (FGFs), epidermal growth factors (EGFs), insulin-like growth factors (IGFs), transforming growth factor beta (TGFb), platelet derived growth factor (PDGFs), hepatocyte growth factor (HGFs), or tumor necrosis factor (TNFs).

[0414] A host cell can also be cultured under conditions that induce stress, e.g., cellular stress, osmotic stress, translational stress, or oncogenic stress. In an embodiment, a host cell expressing a TREM, cultured under conditions that induce stress (e.g., as described herein) results in a fragment of the TREM, e.g., as described herein.

[0415] A host cell can be cultured under nutrient limiting conditions, e.g., the host cell is cultured in media that has a limited amount of one or more nutrients. Examples of nutrients that can be limiting are amino acids, lipids, carbohydrates, hormones, growth factors or vitamins. In an embodiment, a host cell expressing a TREM, cultured in media that has a limited amount of one or more nutrients, e.g., the media is nutrient starved, results in a fragment of the TREM, e.g., as described herein. In an embodiment, a host cell expressing a TREM, cultured in media that has a limited amount of one or more nutrients, e.g., the media is nutrient starved, results in a TREM that is uncharged (e.g. a uTREM).

[0416] A host cell can comprise an immortalized cell, e.g., a cell which expresses one or more enzymes involved in immortalization, e.g., TERT. In an embodiment, a host cell can be propagated indefinitely.

[0417] A host cell can be cultured in suspension or as a monolayer. Host cell cultures can be performed in a cell culture vessel or a bioreactor. Cell culture vessels include a cell culture dish, plate or flask. Exemplary cell culture vessels include 35 mm, 60 mm, 100 mm, or 150 mm dishes, multi-well plates (e.g., 6-well, 12-well, 24-well, 48-well or 96 well plates), or T-25, T-75 or T-160 flasks.

[0418] In an embodiment, a host cell can be cultured in a bioreactor. A bioreactor can be, e.g., a continuous flow batch bioreactor, a perfusion bioreactor, a batch process bioreactor or a fed batch bioreactor. A bioreactor can be maintained under conditions sufficient to express the TREM. The culture conditions can be modulated to optimize yield, purity or structure of the TREM. In an embodiment, a bioreactor comprises at least 1×10.sup.7, 1×10.sup.8, 1×10.sup.9, 1×10.sup.10, 1×10.sup.11, 1×10.sup.12, 1×10.sup.13, or 1×10.sup.14 host cells. In an embodiment, a bioreactor comprises between 1×10.sup.7 to 1×10.sup.14 host cells; between 1×10.sup.7 to 0.5×10.sup.14 host cells; between 1×10.sup.7 to 1×10.sup.13 host cells; between 1×10.sup.7 to 0.5×10.sup.13 host cells; between 1×10.sup.7 to 1×10.sup.12 host cells; between 1×10.sup.7 to 0.5×10.sup.12 host cells; between 1×10.sup.7 to 1×10.sup.11 host cells; between 1×10.sup.7 to 0.5×10.sup.11 host cells; between 1×10.sup.7 to 1×10.sup.10 host cells; between 1×10.sup.7 to 0.5×10.sup.10 host cells; between 1×10.sup.7 to 1×10.sup.9 host cells; between 1×10.sup.7 to 0.5×10.sup.9 host cells; between 1×10.sup.7 to 1×10.sup.8 host cells; between 1×10.sup.7 to 0.5×10.sup.8 host cells; between 0.5×10.sup.8 to 1×10.sup.14 host cells; between 1×10.sup.8 to 1×10.sup.14 host cells; between 0.5×10.sup.9 to 1×10.sup.14 host cells; between 1×10.sup.9 to 1×10.sup.14 host cells; between 0.5×10.sup.10 to 1×10.sup.14 host cells; between 1×10.sup.10 to 1×10.sup.14 host cells; between 0.5×10.sup.11 to 1×10.sup.14 host cells; between 1×10.sup.11 to 1×10.sup.14 host cells; between 0.5×10.sup.12 to 1×10.sup.14 host cells; between 1×10.sup.12 to 1×10.sup.14 host cells; between 0.5×10.sup.13 to 1×10.sup.14 host cells; between 1×10.sup.13 to 1×10.sup.14 host cells; or between 0.5×10.sup.13 to 1×10.sup.14 host cells.

[0419] In an embodiment, a bioreactor comprises at least 1×10.sup.5 host cells/mL, 2×10.sup.5 host cells/mL, 3×10.sup.5 host cells/mL, 4×10.sup.5 host cells/mL, 5×10.sup.5 host cells/mL, 6×10.sup.5 host cells/mL, 7×10.sup.5 host cells/mL, 8×10.sup.5 host cells/mL, 9×10.sup.5 host cells/mL, 1×10.sup.6 host cells/mL, 2×10.sup.6 host cells/mL, 3×10.sup.6 host cells/mL, 4×10.sup.6 host cells/mL, 5×10.sup.6 host cells/mL, 6×10.sup.6 host cells/mL, 7×10.sup.6 host cells/mL, 8×10.sup.6 host cells/mL, 9×10.sup.6 host cells/mL, 1×10.sup.7 host cells/mL, 2×10.sup.7 host cells/mL, 3×10.sup.7 host cells/mL, 4×10.sup.7 host cells/mL, 5×10.sup.7 host cells/mL, 6×10.sup.7 host cells/mL, 7×10.sup.7 host cells/mL, 8×10.sup.7 host cells/mL, 9×10.sup.7 host cells/mL, 1×10.sup.8 host cell/mL, 2×10.sup.8 host cells/mL, 3×10.sup.8 host cells/mL, 4×10.sup.8 host cells/mL, 5×10.sup.8 host cells/mL, 6×10.sup.8 host cells/mL, 7×10.sup.8 host cells/mL, 8×10.sup.8 host cells/mL, 9×10.sup.8 host cells/mL, or 1×10.sup.9 host cells/mL. In an embodiment, a bioreactor comprises between 1×10.sup.5 host cells/mL to 1×10.sup.9 host cells/mL, between 5×10.sup.5 host cells/mL to 1×10.sup.9 host cells/mL, between 1×10.sup.6 host cells/mL to 1×10.sup.9 host cells/mL; between 5×10.sup.6 host cells/mL to 1×10.sup.9 host cells/mL, between 1×10.sup.7 host cells/mL to 1×10.sup.9 host cells/mL, between 5×10.sup.7 host cells/mL to 1×10.sup.9 host cells/mL, between 1×10.sup.8 host cells/mL to 1×10.sup.9 host cells/mL, between 5×10.sup.8 host cells/mL to 1×10.sup.9 host cells/mL, between 1×10.sup.5 host cells/mL to 5×10.sup.8 host cells/mL, between 1×10.sup.5 host cells/mL to 1×10.sup.8 host cells/mL, between 1×10.sup.5 host cells/mL to 5×10.sup.7 host cells/mL, between 1×10.sup.5 host cells/mL to 1×10.sup.7 host cells/mL, between 1×10.sup.5 host cells/mL to 5×10.sup.6 host cells/mL, between 1×10.sup.5 host cells/mL to 1×10.sup.6 host cells/mL, or between 1×10.sup.5 host cells/mL to 5×10.sup.5 host cells/mL.

[0420] In an embodiment, a batch process bioreactor comprises 1×10.sup.6 to 1×10.sup.7 host cells/ml.

[0421] In an embodiment, a batch process bioreactor with a 100 mL volume comprises 1×10.sup.8 to 1×10.sup.9 host cells.

[0422] In an embodiment, a batch process bioreactor with a 100 L volume comprises 1×10.sup.11 to 1×10.sup.12 host cells.

[0423] In an embodiment, a fed batch bioreactor comprises 1×10.sup.7 to 3×10.sup.7 host cells/ml.

[0424] In an embodiment, a fed batch bioreactor with a 100 mL volume comprises 1×10.sup.9 to 3×10.sup.9 host cells.

[0425] In an embodiment, a fed batch bioreactor with a 100 L volume comprises 1×10.sup.12 to 3×10.sup.12 host cells.

[0426] In an embodiment, a perfusion bioreactor comprises 1×10.sup.8 host cells/ml.

[0427] In an embodiment, a perfusion bioreactor with a 100 mL volume comprises 1×10.sup.10 host cells.

[0428] In an embodiment, a perfusion bioreactor with a 100 L volume comprises 1×10.sup.13 host cells.

[0429] In an embodiment, a bioreactor is maintained under conditions that promote growth of the host cell, e.g., at a temperature (e.g., 37° C.) and gas concentration (e.g., 5% CO.sub.2) that is permissive for growth of the host cell.

[0430] For example, in some aspects, a bioreactor unit can perform one or more, or all, of the following: feeding of nutrients and/or carbon sources, injection of suitable gas (e.g., oxygen), inlet and outlet flow of fermentation or cell culture medium, separation of gas and liquid phases, maintenance of temperature, maintenance of oxygen and CO2 levels, maintenance of pH level, agitation (e.g., stirring), and/or cleaning/sterilizing. Exemplary bioreactor units, may contain multiple reactors within the unit, for example the unit can have 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, or 100, or more bioreactors in each unit and/or a facility may contain multiple units having a single or multiple reactors within the facility. Any suitable bioreactor diameter can be used.

[0431] In an embodiment, the bioreactor can have a volume between about 100 mL and about 100 L. Non-limiting examples include a volume of 100 mL, 250 mL, 500 mL, 750 mL, 1 liter, 2 liters, 3 liters, 4 liters, 5 liters, 6 liters, 7 liters, 8 liters, 9 liters, 10 liters, 15 liters, 20 liters, 25 liters, 30 liters, 40 liters, 50 liters, 60 liters, 70 liters, 80 liters, 90 liters, 100 liters. Additionally, suitable reactors can be multi-use, single-use, disposable, or non-disposable and can be formed of any suitable material including metal alloys such as stainless steel (e.g., 316L or any other suitable stainless steel) and Inconel, plastics, and/or glass. In some embodiments, suitable reactors can be round, e.g., cylindrical. In some embodiments, suitable reactors can be square, e.g., rectangular. Square reactors may in some cases provide benefits over round reactors such as ease of use (e.g., loading and setup by skilled persons), greater mixing and homogeneity of reactor contents, and lower floor footprint.

Method of Modifying Host Cells

[0432] A host cell can be modified to optimize the production of a TREM, e.g., to have optimized TREM yield, purity, structure (e.g., folding), or stability. In an embodiment, a host cell can be modified (e.g., using a method described herein), to increase or decrease the expression of a desired molecule, e.g., gene, which optimizes production of the TREM, e.g., optimizes yield, purity, structure or stability of the TREM. In an embodiment, a host cell can be epigenetically modified, e.g., using a method described herein, to increase or decrease the expression of a desired gene, which optimizes production.

[0433] In an embodiment, a host cell can be modified to increase or decrease the expression of an oncogene (e.g., as described herein), a tumor suppressor (e.g., as described herein) or a molecule involved in tRNA or TREM modulation (e.g., a gene involved in tRNA or TREM transcription, processing, modification, stability or folding). Exemplary oncogenes include Myc (e.g., c-Myc, N-Myc or L-Myc), c-Jun, Wnt, or RAS. Exemplary tumor suppressors include p53 or Rb. Exemplary molecules involved in tRNA or TREM modulation include: RNA Polymerase III (Pol III) and Pol III accessory molecules (e.g., TFIIIB); Maf1, Trm1, Mck1 or Kns 1; enzymes involved in tRNA or TREM modification, e.g., genes listed in Table 2; or molecules with nuclease activity, e.g., or one or more of Dicer, Angiogenin, RNaseA, RNaseP, RNaseZ, Rny1 or PrrC.

[0434] In an embodiment, a host cell can be modified by: transfection (e.g., transient transfection or stable transfection); transduction (e.g., viral transduction, e.g., lentiviral, adenoviral or retroviral transduction); electroporation; lipid-based delivery of an agent (e.g., liposomes), nanoparticle based delivery of an agent; or other methods known in the art.

[0435] In an embodiment, a host cell can be modified to increase the expression of, e.g., overexpress, a desired molecule, e.g., a gene (e.g., an oncogene, or a gene involved in tRNA or TREM modulation (e.g., a gene encoding an enzyme listed in Table 2, or a gene encoding an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., or one or more of Dicer, Angiogenin, RNaseA, RNaseP, RNaseZ, Rny1 or PrrC. Exemplary methods of increasing the expression of a gene include: (a) contacting the host cell with a nucleic acid (e.g., DNA, or RNA) encoding the gene; (b) contacting the host cell with a peptide that expresses the target protein; (c) contacting the host cell with a molecule (e.g., a small RNA (e.g., a micro RNA, or a small interfering RNA) or a low molecular weight compound) that modulates, e.g., increases the expression of the target gene; or (d) contacting the host cell with a gene editing moiety (e.g., a zinc finger nuclease (ZFN) or a Cas9/CRISPR molecule) that inhibits (e.g., mutates or knocks-out) the expression of a negative regulator of the target gene. In an embodiment, a nucleic acid encoding the gene, or a plasmid containing a nucleic acid encoding the gene can be introduced into the host cell by transfection or electroporation. In an embodiment, a nucleic acid encoding a gene can be introduced into the host cell by contacting the host cell with a virus (e.g., a lentivirus, adenovirus or retrovirus) expressing the gene.

[0436] In an embodiment, a host cell can be modified to decrease the expression of, e.g., minimize the expression, of a desired molecule, e.g., a gene (e.g., a tumor suppressor, or a gene involved in tRNA or TREM modulation). Exemplary methods of decreasing the expression of a gene include: (a) contacting the host cell with a nucleic acid (e.g., DNA, or RNA) encoding an inhibitor of the gene (e.g., a dominant negative variant or a negative regulator of the gene or protein encoded by the gene); (b) contacting the host cell with a peptide that inhibits the target protein; (c) contacting the host cell with a molecule (e.g., a small RNA (e.g., a micro RNA, or a small interfering RNA) or a low molecular weight compound) that modulates, e.g., inhibits the expression of the target gene; or (d) contacting the host cell with a gene editing moiety (e.g., a zinc finger nuclease (ZFN) or a Cas9/CRISPR molecule) that inhibits (e.g., mutates or knocks-out) the expression of the target gene. In an embodiment, a nucleic acid encoding an inhibitor of the gene, or a plasmid containing a nucleic acid encoding an inhibitor of the gene can be introduced into the host cell by transfection or electroporation. In an embodiment, a nucleic acid encoding an inhibitor of the gene can be introduced into the host cell by contacting the host cell with a virus (e.g., a lentivirus, adenovirus or retrovirus) expressing the inhibitor of the gene.

[0437] In an embodiment, a host cell (e.g., a host cell described herein) is modified (e.g., by transfection with a nucleic acid), to express, e.g., overexpress, an oncogene, e.g., an oncogene described herein, e.g., c-Myc.

[0438] In an embodiment, a host cell (e.g., a host cell described herein) is modified (e.g., by transfection with a nucleic acid), to repress, e.g., downregulate, expression of a tumor suppressor, e.g., a tumor suppressor described herein, e.g., p53 or Rb.

[0439] In an embodiment, a host cell (e.g., a HEK293T cell) is modified (e.g., using a CRISPR/Cas9 molecule) to inhibit, e.g., knockout, expression of a gene that modulates a tRNA or TREM, e.g., Maf1. In an embodiment, a host cell (e.g., a HEK293T cell) is modified to overexpress a gene that modulates a tRNA or TREM, e.g., Trm1.

[0440] In an embodiment, a host cell (e.g., a HEK293T cell) is modified to overexpress a gene that modulates a tRNA or TREM, e.g., Trm1, and to overexpress an oncogene, e.g., an oncogene described herein, e.g., c-Myc.

TREM

[0441] A “tRNA-based effector molecule” or “TREM” refers to an RNA molecule comprising one or more of the properties described herein. A TREM can be charged with an amino acid, e.g., a cognate amino acid; charged with a non-cognate amino acid (e.g., a mischarged TREM (mTREM); or not charged with an amino acid, e.g., an uncharged TREM (uTREM).

[0442] In an embodiment, a TREM comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM comprises an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0443] In an embodiment, a TREM comprises at least 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., at least 30 consecutive nucleotides of an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM comprises at least 30 consecutive nucleotides of an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM comprises at least 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

TABLE-US-00001 TABLE 1 List of tRNA sequences SEQ ID NO tRNA name tRNA sequence 1 Ala_AGC_chr6: 28763741-28763812 (−) GGGGGTATAGCTCAGTGGTAGAGCGCGTGCTTAGCATGCACGAGGTCC TGGGTTCGATCCCCAGTACCTCCA 2 Ala_AGC_chr6: 26687485-26687557 (+) GGGGAATTAGCTCAAGTGGTAGAGCGCTTGCTTAGCACGCAAGAGGTA GTGGGATCGATGCCCACATTCTCCA 3 Ala_AGC_chr6: 26572092-26572164 (−) GGGGAATTAGCTCAAATGGTAGAGCGCTCGCTTAGCATGCGAGAGGTA GCGGGATCGATGCCCGCATTCTCCA 4 Ala_AGC_chr6: 26682715-26682787 (+) GGGGAATTAGCTCAAGTGGTAGAGCGCTTGCTTAGCATGCAAGAGGTA GTGGGATCGATGCCCACATTCTCCA 5 Ala_AGC_chr6: 26705606-26705678 (+) GGGGAATTAGCTCAAGCGGTAGAGCGCTTGCTTAGCATGCAAGAGGTA GTGGGATCGATGCCCACATTCTCCA 6 Ala_AGC_chr6: 26673590-26673662 (+) GGGGAATTAGCTCAAGTGGTAGAGCGCTTGCTTAGCATGCAAGAGGTA GTGGGATCAATGCCCACATTCTCCA 7 Ala_AGC_chr14: 89445442-89445514 (+) GGGGAATTAGCTCAAGTGGTAGAGCGCTCGCTTAGCATGCGAGAGGTA GTGGGATCGATGCCCGCATTCTCCA 8 Ala_AGC_chr6: 58196623-58196695 (−) GGGGAATTAGCCCAAGTGGTAGAGCGCTTGCTTAGCATGCAAGAGGTA GTGGGATCGATGCCCACATTCTCCA 9 Ala_AGC_chr6: 28806221-28806292 (−) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGCACGAGGCCC CGGGTTCAATCCCCGGCACCTCCA 10 Ala_AGC_chr6: 28574933-28575004 (+) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGTACGAGGTCC CGGGTTCAATCCCCGGCACCTCCA 11 Ala_AGC_chr6: 28626014-28626085 (−) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTAGCATGCATGAGGTCC CGGGTTCGATCCCCAGCATCTCCA 12 Ala_AGC_chr6: 28678366-28678437 (+) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGCACGAGGCCC TGGGTTCAATCCCCAGCACCTCCA 13 Ala_AGC_chr6: 28779849-28779920 (−) GGGGGTATAGCTCAGCGGTAGAGCGCGTGCTTAGCATGCACGAGGTCC TGGGTTCAATCCCCAATACCTCCA 14 Ala_AGC_chr6: 28687481-28687552 (+) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGCACGAGGCCC CGGGTTCAATCCCTGGCACCTCCA 15 Ala_AGC_chr2: 27274082-27274154 (+) GGGGGATTAGCTCAAATGGTAGAGCGCTCGCTTAGCATGCGAGAGGTA GCGGGATCGATGCCCGCATCCTCCA 16 Ala_AGC_chr6: 26730737-26730809 (+) GGGGAATTAGCTCAGGCGGTAGAGCGCTCGCTTAGCATGCGAGAGGTA GCGGGATCGACGCCCGCATTCTCCA 17 Ala_CGC_chr6: 26553731-26553802 (+) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTCGCATGTATGAGGTCC CGGGTTCGATCCCCGGCATCTCCA 18 Ala_CGC_chr6: 28641613-28641684 (−) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTCGCATGTATGAGGCCC CGGGTTCGATCCCCGGCATCTCCA 19 Ala_CGC_chr2: 157257281-157257352 (+) GGGGATGTAGCTCAGTGGTAGAGCGCGCGCTTCGCATGTGTGAGGTCC CGGGTTCAATCCCCGGCATCTCCA 20 Ala_CGC_chr6: 28697092-28697163 (+) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTCGCATGTACGAGGCCC CGGGTTCGACCCCCGGCTCCTCCA 21 Ala_TGC_chr6: 28757547-28757618 (−) GGGGGTGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGTCC CGGGTTCGATCCCCGGCACCTCCA 22 Ala_TGC_chr6: 28611222-28611293 (+) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGTCC CGGGTTCGATCCCCGGCATCTCCA 23 Ala_TGC_chr5: 180633868-180633939 (+) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGCCC CGGGTTCGATCCCCGGCATCTCCA 24 Ala_TGC_chr12: 125424512-125424583 (+) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTTGCACGTATGAGGCCC CGGGTTCAATCCCCGGCATCTCCA 25 Ala_TGC_chr6: 28785012-28785083 (−) GGGGGTGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGCCT CGGGTTCGATCCCCGACACCTCCA 26 Ala_TGC_chr6: 28726141-28726212 (−) GGGGGTGTAGCTCAGTGGTAGAGCACATGCTTTGCATGTGTGAGGCCC CGGGTTCGATCCCCGGCACCTCCA 27 Ala_TGC_chr6: 28770577-28770647 (−) GGGGGTGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGCCT CGGTTCGATCCCCGACACCTCCA 28 Arg_ACG_chr6: 26328368-26328440 (+) GGGCCAGTGGCGCAATGGATAACGCGTCTGACTACGGATCAGAAGATT CCAGGTTCGACTCCTGGCTGGCTCG 29 Arg_ACG_chr3: 45730491-45730563 (−) GGGCCAGTGGCGCAATGGATAACGCGTCTGACTACGGATCAGAAGATT CTAGGTTCGACTCCTGGCTGGCTCG 30 Arg_CCG_chr6: 28710729-28710801 (−) GGCCGCGTGGCCTAATGGATAAGGCGTCTGATTCCGGATCAGAAGATT GAGGGTTCGAGTCCCTTCGTGGTCG 31 Arg_CCG_chr17: 66016013-66016085 (−) GACCCAGTGGCCTAATGGATAAGGCATCAGCCTCCGGAGCTGGGGATT GTGGGTTCGAGTCCCATCTGGGTCG 32 Arg_CCT_chr17: 73030001-73030073 (+) GCCCCAGTGGCCTAATGGATAAGGCACTGGCCTCCTAAGCCAGGGATT GTGGGTTCGAGTCCCACCTGGGGTA 33 Arg_CCT_chr17: 73030526-73030598 (−) GCCCCAGTGGCCTAATGGATAAGGCACTGGCCTCCTAAGCCAGGGATT GTGGGTTCGAGTCCCACCTGGGGTG 34 Arg_CCT_chr16: 3202901-3202973 (+) GCCCCGGTGGCCTAATGGATAAGGCATTGGCCTCCTAAGCCAGGGATT GTGGGTTCGAGTCCCACCCGGGGTA 35 Arg_CCT_chr7: 139025446-139025518 (+) GCCCCAGTGGCCTAATGGATAAGGCATTGGCCTCCTAAGCCAGGGATT GTGGGTTCGAGTCCCATCTGGGGTG 36 Arg_CCT_chr16: 3243918-3243990 (+) GCCCCAGTGGCCTGATGGATAAGGTACTGGCCTCCTAAGCCAGGGATT GTGGGTTCGAGTTCCACCTGGGGTA 37 Arg_TCG_chr15: 89878304-89878376 (+) GGCCGCGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGATT GCAGGTTCGAGTCCTGCCGCGGTCG 38 Arg_TCG_chr6: 26323046-26323118 (+) GACCACGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGATT GAGGGTTCGAATCCCTCCGTGGTTA 39 Arg_TCG_chr17: 73031208-73031280 (+) GACCGCGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGATT GAGGGTTCGAGTCCCTTCGTGGTCG 40 Arg_TCG_chr6: 26299905-26299977 (+) GACCACGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGATT GAGGGTTCGAATCCCTTCGTGGTTA 41 Arg_TCG_chr6: 28510891-28510963 (−) GACCACGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGATT GAGGGTTCGAATCCCTTCGTGGTTG 42 Arg_TCG_chr9: 112960803-112960875 (+) GGCCGTGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAAAAGATT GCAGGTTTGAGTTCTGCCACGGTCG 43 Arg_TCT_chr1: 94313129-94313213 (+) GGCTCCGTGGCGCAATGGATAGCGCATTGGACTTCTAGAGGCTGAAGG CATTCAAAGGTTCCGGGTTCGAGTCCCGGCGGAGTCG 44 Arg_TCT_chr17: 8024243-8024330 (+) GGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGTGACGAATAG AGCAATTCAAAGGTTGTGGGTTCGAATCCCACCAGAGTCG 45 Arg_TCT_chr9: 131102355-131102445 (−) GGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGCTGAGCCTAG TGTGGTCATTCAAAGGTTGTGGGTTCGAGTCCCACCAGAGTCG 46 Arg_TCT_chr11: 59318767-59318852 (+) GGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGATAGTTAGAG AAATTCAAAGGTTGTGGGTTCGAGTCCCACCAGAGTCG 47 Arg_TCT_chr1: 159111401-159111474 (−) GTCTCTGTGGCGCAATGGACGAGCGCGCTGGACTTCTAATCCAGAGGT TCCGGGTTCGAGTCCCGGCAGAGATG 48 Arg_TCT_chr6: 27529963-27530049 (+) GGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGCCTAAATCAA GAGATTCAAAGGTTGCGGGTTCGAGTCCCTCCAGAGTCG 49 Asn_GTT_chr1: 161510031-161510104 (+) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGGT TGGTGGTTCGATCCCACCCAGGGACG 50 Asn_GTT_chr1: 143879832-143879905 (−) GTCTCTGTGGCGCAATCGGCTAGCGCGTTTGGCTGTTAACTAAAAGGTT GGCGGTTCGAACCCACCCAGAGGCG 51 Asn_GTT_chr1: 144301611-144301684 (+) GTCTCTGTGGTGCAATCGGTTAGCGCGTTCCGCTGTTAACCGAAAGCTT GGTGGTTCGAGCCCACCCAGGGATG 52 Asn_GTT_chr1: 149326272-149326345 (−) GTCTCTGTGGCGCAATCGGCTAGCGCGTTTGGCTGTTAACTAAAAAGTT GGTGGTTCGAACACACCCAGAGGCG 53 Asn_GTT_chr1: 148248115-148248188 (+) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCACCCAGGGACG 54 Asn_GTT_chr1: 148598314-148598387 (−) GTCTCTGTGGCGCAATCGGTTAGCGCATTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCACCCAGGGACG 55 Asn_GTT_chr1: 17216172-17216245 (+) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGAT TGGTGGTTCGAGCCCACCCAGGGACG 56 Asn_GTT_chr1: 16847080-16847153 (−) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACTGAAAGGTT GGTGGTTCGAGCCCACCCAGGGACG 57 Asn_GTT_chr1: 149230570-149230643 (−) GTCTCTGTGGCGCAATGGGTTAGCGCGTTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCATCCAGGGACG 58 Asn_GTT_chr1: 148000805-148000878 (+) GTCTCTGTGGCGTAGTCGGTTAGCGCGTTCGGCTGTTAACCGAAAAGTT GGTGGTTCGAGCCCACCCAGGAACG 59 Asn_GTT_chr1: 149711798-149711871 (−) GTCTCTGTGGCGCAATCGGCTAGCGCGTTTGGCTGTTAACTAAAAGGTT GGTGGTTCGAACCCACCCAGAGGCG 60 Asn_GTT_chr1: 145979034-145979107 (−) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACTGAAAGGTT AGTGGTTCGAGCCCACCCGGGGACG 61 Asp_GTC_chr12: 98897281-98897352 (+) TCCTCGTTAGTATAGTGGTTAGTATCCCCGCCTGTCACGCGGGAGACCG GGGTTCAATTCCCCGACGGGGAG 62 Asp_GTC_chr1: 161410615-161410686 (−) TCCTCGTTAGTATAGTGGTGAGTATCCCCGCCTGTCACGCGGGAGACC GGGGTTCGATTCCCCGACGGGGAG 63 Asp_GTC_chr6: 27551236-27551307 (−) TCCTCGTTAGTATAGTGGTGAGTGTCCCCGTCTGTCACGCGGGAGACC GGGGTTCGATTCCCCGACGGGGAG 64 Cys_GCA_chr7: 149007281-149007352 (+) GGGGGCATAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCAGGTGCCCCCT 65 Cys_GCA_chr7: 149074601-149074672 (−) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCAGGTGCCCCCC 66 Cys_GCA_chr7: 149112229-149112300 (−) GGGGGTATAGCTTAGCGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGGTGCCCCCT 67 Cys_GCA_chr7: 149344046-149344117 (−) GGGGGTATAGCTTAGGGGTAGAGCATTTGACTGCAGATCAAAAGGTCC CTGGTTCAAATCCAGGTGCCCCTT 68 Cys_GCA_chr7: 149052766-149052837 (−) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCAGTTCAAATCTGGGTGCCCCCT 69 Cys_GCA_chr17: 37017937-37018008 (−) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAAGTCC CCGGTTCAAATCCGGGTGCCCCCT 70 Cys_GCA_chr7: 149281816-149281887 (+) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCT CTGGTTCAAATCCAGGTGCCCCCT 71 Cys_GCA_chr7: 149243631-149243702 (+) GGGGGTATAGCTCAGGGGTAGAGCACTTGACTGCAGATCAAGAAGTCC TTGGTTCAAATCCAGGTGCCCCCT 72 Cys_GCA_chr7: 149388272-149388343 (−) GGGGATATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGGTGCCCCCC 73 Cys_GCA_chr7: 149072850-149072921 (−) GGGGGTATAGTTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCAGGTGCCCCCT 74 Cys_GCA_chr7: 149310156-149310227 (−) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAAATCAAGAGGTCC CTGATTCAAATCCAGGTGCCCCCT 75 Cys_GCA_chr4: 124430005-124430076 (−) GGGGGTATAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGGTGCCCCCT 76 Cys_GCA_chr7: 149295046-149295117 (+) GGGCGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCAGTTCAAATCTGGGTGCCCCCT 77 Cys_GCA_chr7: 149361915-149361986 (+) GGGGGTATAGCTCACAGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCTGGGTGCCCCCT 78 Cys_GCA_chr7: 149253802-149253871 (+) GGGCGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCAGTTCAAATCTGGGTGCCCA 79 Cys_GCA_chr7: 149292305-149292376 (−) GGGGGTATAGCTCACAGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGTTACTCCCT 80 Cys_GCA_chr7: 149286164-149286235 (−) GGGGGTATAGCTCAGGGGTAGAGCACTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCAGGTGCCCCCT 81 Cys_GCA_chr17: 37025545-37025616 (−) GGGGGTATAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCGGGTGCCCCCT 82 Cys_GCA_chr15: 80036997-80037069 (+) GGGGGTATAGCTCAGTGGGTAGAGCATTTGACTGCAGATCAAGAGGTC CCCGGTTCAAATCCGGGTGCCCCCT 83 Cys_GCA_chr3: 131947944-131948015 (−) GGGGGTGTAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCAGGTGCCCCCT 84 Cys_GCA_chr1: 93981834-93981906 (−) GGGGGTATAGCTCAGGTGGTAGAGCATTTGACTGCAGATCAAGAGGTC CCCGGTTCAAATCCGGGTGCCCCCT 85 Cys_GCA_chr14: 73429679-73429750 (+) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGGTGCCCCCT 86 Cys_GCA_chr3: 131950642-131950713 (−) GGGGGTATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCAGGTGCCCCCT 87 Gln_CTG_chr6: 18836402-18836473 (+) GGTTCCATGGTGTAATGGTTAGCACTCTGGACTCTGAATCCAGCGATCC GAGTTCAAATCTCGGTGGAACCT 88 Gln_CTG_chr6: 27515531-27515602 (−) GGTTCCATGGTGTAATGGTTAGCACTCTGGACTCTGAATCCAGCGATCC GAGTTCAAGTCTCGGTGGAACCT 89 Gln_CTG_chr1: 145963304-145963375 (+) GGTTCCATGGTGTAATGGTGAGCACTCTGGACTCTGAATCCAGCGATC CGAGTTCGAGTCTCGGTGGAACCT 90 Gln_CTG_chr1: 147737382-147737453 (−) GGTTCCATGGTGTAATGGTAAGCACTCTGGACTCTGAATCCAGCGATC CGAGTTCGAGTCTCGGTGGAACCT 91 Gln_CTG_chr6: 27263212-27263283 (+) GGTTCCATGGTGTAATGGTTAGCACTCTGGACTCTGAATCCGGTAATCC GAGTTCAAATCTCGGTGGAACCT 92 Gln_CTG_chr6: 27759135-27759206 (−) GGCCCCATGGTGTAATGGTCAGCACTCTGGACTCTGAATCCAGCGATC CGAGTTCAAATCTCGGTGGGACCC 93 Gln_CTG_chr1: 147800937-147801008 (+) GGTTCCATGGTGTAATGGTAAGCACTCTGGACTCTGAATCCAGCCATCT GAGTTCGAGTCTCTGTGGAACCT 94 Gln_TTG_chr17: 47269890-47269961 (+) GGTCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCGATCC GAGTTCAAATCTCGGTGGGACCT 95 Gln_TTG_chr6: 28557156-28557227 (+) GGTCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCAATCC GAGTTCGAATCTCGGTGGGACCT 96 Gln_TTG_chr6: 26311424-26311495 (−) GGCCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCGATC CGAGTTCAAATCTCGGTGGGACCT 97 Gln_TTG_chr6: 145503859-145503930 (+) GGTCCCATGGTGTAATGGTTAGCACTCTGGGCTTTGAATCCAGCAATCC GAGTTCGAATCTTGGTGGGACCT 98 Glu_CTC_chr1: 145399233-145399304 (−) TCCCTGGTGGTCTAGTGGTTAGGATTCGGCGCTCTCACCGCCGCGGCCC GGGTTCGATTCCCGGTCAGGGAA 99 Glu_CTC_chr1: 249168447-249168518 (+) TCCCTGGTGGTCTAGTGGTTAGGATTCGGCGCTCTCACCGCCGCGGCCC GGGTTCGATTCCCGGTCAGGAAA 100 Glu_TTC_chr2: 131094701-131094772 (−) TCCCATATGGTCTAGCGGTTAGGATTCCTGGTTTTCACCCAGGTGGCCC GGGTTCGACTCCCGGTATGGGAA 101 Glu_TTC_chr13 :45492062-45492133 (−) TCCCACATGGTCTAGCGGTTAGGATTCCTGGTTTTCACCCAGGCGGCCC GGGTTCGACTCCCGGTGTGGGAA 102 Glu_TTC_chr1: 17199078-17199149 (+) TCCCTGGTGGTCTAGTGGCTAGGATTCGGCGCTTTCACCGCCGCGGCCC GGGTTCGATTCCCGGCCAGGGAA 103 Glu_TTC_chr1: 16861774-16861845 (−) TCCCTGGTGGTCTAGTGGCTAGGATTCGGCGCTTTCACCGCCGCGGCCC GGGTTCGATTCCCGGTCAGGGAA 104 Gly_CCC_chr1 :16872434-16872504 (−) GCATTGGTGGTTCAGTGGTAGAATTCTCGCCTCCCACGCGGGAGACCC GGGTTCAATTCCCGGCCAATGCA 105 Gly_CCC_chr2:70476123-70476193 (−) GCGCCGCTGGTGTAGTGGTATCATGCAAGATTCCCATTCTTGCGACCCG GGTTCGATTCCCGGGCGGCGCA 106 Gly_CCC_chr17 : 19764175-19764245 (+) GCATTGGTGGTTCAATGGTAGAATTCTCGCCTCCCACGCAGGAGACCC AGGTTCGATTCCTGGCCAATGCA 107 Gly_GCC_chr1: 161413094-161413164 (+) GCATGGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCCC GGGTTCGATTCCCGGCCCATGCA 108 Gly_GCC_chr1: 161493637-161493707 (−) GCATTGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCCC GGGTTCGATTCCCGGCCAATGCA 109 Gly_GCC_chr16: 70812114-70812184 (−) GCATTGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCCC GGGTTTGATTCCCGGCCAGTGCA 110 Gly_GCC_chr1: 161450356-161450426 (+) GCATAGGTGGTTCAGTGGTAGAATTCTTGCCTGCCACGCAGGAGGCCC AGGTTTGATTCCTGGCCCATGCA 111 Gly_GCC_chr16:70822597-70822667 (+) GCATTGGTGGTTCAGTGGTAGAATTCTCGCCTGCCATGCGGGCGGCCG GGCTTCGATTCCTGGCCAATGCA 112 Gly_TCC_chr19: 4724082-4724153 (+) GCGTTGGTGGTATAGTGGTTAGCATAGCTGCCTTCCAAGCAGTTGACC CGGGTTCGATTCCCGGCCAACGCA 113 Gly_TCC_chr1: 145397864-145397935 (−) GCGTTGGTGGTATAGTGGTGAGCATAGCTGCCTTCCAAGCAGTTGACC CGGGTTCGATTCCCGGCCAACGCA 114 Gly_TCC_chr17: 8124866-8124937 (+) GCGTTGGTGGTATAGTGGTAAGCATAGCTGCCTTCCAAGCAGTTGACC CGGGTTCGATTCCCGGCCAACGCA 115 Gly_TCC_chr1: 161409961-161410032 (−) GCGTTGGTGGTATAGTGGTGAGCATAGTTGCCTTCCAAGCAGTTGACC CGGGCTCGATTCCCGCCCAACGCA 116 His_GTG_chr1: 145396881-145396952 (−) GCCGTGATCGTATAGTGGTTAGTACTCTGCGTTGTGGCCGCAGCAACCT CGGTTCGAATCCGAGTCACGGCA 117 His_GTG_chr1: 149155828-149155899 (−) GCCATGATCGTATAGTGGTTAGTACTCTGCGCTGTGGCCGCAGCAACC TCGGTTCGAATCCGAGTCACGGCA 118 Ile_AAT_chr6: 58149254-58149327 (+) GGCCGGTTAGCTCAGTTGGTTAGAGCGTGGCGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACGGGCCA 119 Ile_AAT_chr6: 27655967-27656040 (+) GGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACTGGCCA 120 Ile_AAT_chr6: 27242990-27243063 (−) GGCTGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACTGGCCA 121 Ile_AAT_chr17: 8130309-8130382 (−) GGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGAACCCCGTACGGGCCA 122 Ile_AAT_chr6: 26554350-26554423 (+) GGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACGGGCCA 123 Ile_AAT_chr6: 26745255-26745328 (−) GGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCTAAGGT CGCGGGTTCGATCCCCGTACTGGCCA 124 Ile_AAT_chr6: 26721221-26721294 (−) GGCCGGTTAGCTCAGTTGGTCAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACGGGCCA 125 Ile_AAT_chr6: 27636362-27636435 (+) GGCCGGTTAGCTCAGTCGGCTAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACGGGCCA 126 Ile_AAT_chr6: 27241739-27241812 (+) GGCTGGTTAGTTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGGT CGTGGGTTCGATCCCCATATCGGCCA 127 Ile_GAT_chrX: 3756418-3756491 (−) GGCCGGTTAGCTCAGTTGGTAAGAGCGTGGTGCTGATAACACCAAGGT CGCGGGCTCGACTCCCGCACCGGCCA 128 Ile_TAT_chr19: 39902808-39902900 (−) GCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATATGACAGTGCG AGCGGAGCAATGCCGAGGTTGTGAGTTCGATCCTCACCTGGAGCA 129 Ile_TAT_chr2: 43037676-43037768 (+) GCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATACAGCAGTACA TGCAGAGCAATGCCGAGGTTGTGAGTTCGAGCCTCACCTGGAGCA 130 Ile_TAT_chr6: 26988125-26988218 (+) GCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATATGGCAGTATG TGTGCGAGTGATGCCGAGGTTGTGAGTTCGAGCCTCACCTGGAGCA 131 Ile_TAT_chr6: 27599200-27599293 (+) GCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATACAACAGTATA TGTGCGGGTGATGCCGAGGTTGTGAGTTCGAGCCTCACCTGGAGCA 132 Ile_TAT_chr6: 28505367-28505460 (+) GCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATAAGACAGTGCA CCTGTGAGCAATGCCGAGGTTGTGAGTTCAAGCCTCACCTGGAGCA 133 Leu_AAG_chr5: 180524474-180524555 (−) GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTAAGGCTCCAGTCTC TTCGGAGGCGTGGGTTCGAATCCCACCGCTGCCA 134 Leu_AAG_chr5: 180614701-180614782 (+) GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTAAGGCTCCAGTCTC TTCGGGGGCGTGGGTTCGAATCCCACCGCTGCCA 135 Leu_AAG_chr6: 28956779-28956860 (+) GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTAAGGCTCCAGTCTC TTCGGGGGCGTGGGTTCAAATCCCACCGCTGCCA 136 Leu_AAG_chr6: 28446400-28446481 (−) GGTAGCGTGGCCGAGTGGTCTAAGACGCTGGATTAAGGCTCCAGTCTC TTCGGGGGCGTGGGTTTGAATCCCACCGCTGCCA 137 Leu_CAA_chr6: 28864000-28864105 (−) GTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGCTAAGCTTCC TCCGCGGTGGGGATTCTGGTCTCCAATGGAGGCGTGGGTTCGAATCCC 138 Leu_CAA_chr6: 28908830-28908934 (+) GTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGCTTGGCTTCC TCGTGTTGAGGATTCTGGTCTCCAATGGAGGCGTGGGTTCGAATCCCA 139 Leu_CAA_chr6: 27573417-27573524 (−) GTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGCTTACTGCTT CCTGTGTTCGGGTCTTCTGGTCTCCGTATGGAGGCGTGGGTTCGAATCC 140 Leu_CAA_chr6: 27570348-27570454 (−) GTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGTTGCTACTTC CCAGGTTTGGGGCTTCTGGTCTCCGCATGGAGGCGTGGGTTCGAATCC 141 Leu_CAA_chr1: 249168054-249168159 (+) GTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGGTAAGCACCT TGCCTGCGGGCTTTCTGGTCTCCGGATGGAGGCGTGGGTTCGAATCCC 142 Leu_CAA_chr11: 9296790-9296863 (+) GCCTCCTTAGTGCAGTAGGTAGCGCATCAGTCTCAAAATCTGAATGGT CCTGAGTTCAAGCCTCAGAGGGGGCA 143 Leu_CAA_chr1: 161581736-161581819 (−) GTCAGGATGGCCGAGCAGTCTTAAGGCGCTGCGTTCAAATCGCACCCT CCGCTGGAGGCGTGGGTTCGAATCCCACTTTTGACA 144 Leu_CAG_chr1: 161411323-161411405 (+) GTCAGGATGGCCGAGCGGTCTAAGGCGCTGCGTTCAGGTCGCAGTCTC CCCTGGAGGCGTGGGTTCGAATCCCACTCCTGACA 145 Leu_CAG_chr16: 57333863-57333945 (+) GTCAGGATGGCCGAGCGGTCTAAGGCGCTGCGTTCAGGTCGCAGTCTC CCCTGGAGGCGTGGGTTCGAATCCCACTTCTGACA 146 Leu_TAA_chr6: 144537684-144537766 (+) ACCAGGATGGCCGAGTGGTTAAGGCGTTGGACTTAAGATCCAATGGAC ATATGTCCGCGTGGGTTCGAACCCCACTCCTGGTA 147 Leu_TAA_chr6: 27688898-27688980 (−) ACCGGGATGGCCGAGTGGTTAAGGCGTTGGACTTAAGATCCAATGGGC TGGTGCCCGCGTGGGTTCGAACCCCACTCTCGGTA 148 Leu_TAA_chr11: 59319228-59319310 (+) ACCAGAATGGCCGAGTGGTTAAGGCGTTGGACTTAAGATCCAATGGAT TCATATCCGCGTGGGTTCGAACCCCACTTCTGGTA 149 Leu_TAA_chr6: 27198334-27198416 (−) ACCGGGATGGCTGAGTGGTTAAGGCGTTGGACTTAAGATCCAATGGAC AGGTGTCCGCGTGGGTTCGAGCCCCACTCCCGGTA 150 Leu_TAG_chr17: 8023632-8023713 (−) GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTTAGGCTCCAGTCTC TTCGGAGGCGTGGGTTCGAATCCCACCGCTGCCA 151 Leu_TAG_chr14: 21093529-21093610 (+) GGTAGTGTGGCCGAGCGGTCTAAGGCGCTGGATTTAGGCTCCAGTCTC TTCGGGGGCGTGGGTTCGAATCCCACCACTGCCA 152 Leu_TAG_chr16: 22207032-22207113 (−) GGTAGCGTGGCCGAGTGGTCTAAGGCGCTGGATTTAGGCTCCAGTCAT TTCGATGGCGTGGGTTCGAATCCCACCGCTGCCA 153 Lys_CTT_chr14: 58706613-58706685 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGGGACTCTTAATCCCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCG 154 Lys_CTT_chr19: 36066750-36066822 (+) GCCCAGCTAGCTCAGTCGGTAGAGCATAAGACTCTTAATCTCAGGGTT GTGGATTCGTGCCCCATGCTGGGTG 155 Lys_CTT_chr19: 52425393-52425466 (−) GCAGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGTCAT GGGTTCGTGCCCCATGTTGGGTGCCA 156 Lys_CTT_chr1: 145395522-145395594 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCG 157 Lys_CTT_chr16: 3207406-3207478 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGAGACCCTTAATCTCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCG 158 Lys_CTT_chr16: 3241501-3241573 (+) GCCCGGCTAGCTCAGTCGGTAGAGCATGGGACTCTTAATCTCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCG 159 Lys_CTT_chr16: 3230555-3230627 (−) GCCCGGCTAGCTCAGTCGATAGAGCATGAGACTCTTAATCTCAGGGTC GTGGGTTCGAGCCGCACGTTGGGCG 160 Lys_CTT_chr1: 55423542-55423614 (−) GCCCAGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGTC ATGGGTTTGAGCCCCACGTTTGGTG 161 Lys_CTT_chr16: 3214939-3215011 (+) GCCTGGCTAGCTCAGTCGGCAAAGCATGAGACTCTTAATCTCAGGGTC GTGGGCTCGAGCTCCATGTTGGGCG 162 Lys_CTT_chr5: 26198539-26198611 (−) GCCCGACTACCTCAGTCGGTGGAGCATGGGACTCTTCATCCCAGGGTT GTGGGTTCGAGCCCCACATTGGGCA 163 Lys_TTT_chr16: 73512216-73512288 (−) GCCTGGATAGCTCAGTTGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTTCAGGCA 164 Lys_TTT_chr12: 27843306-27843378 (+) ACCCAGATAGCTCAGTCAGTAGAGCATCAGACTTTTAATCTGAGGGTC CAAGGTTCATGTCCCTTTTTGGGTG 165 Lys_TTT_chr11: 122430655-122430727 (+) GCCTGGATAGCTCAGTTGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTTCAGGCG 166 Lys_TTT_chr1: 204475655-204475727 (+) GCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTTCGGGCG 167 Lys_TTT_chr6: 27559593-27559665 (−) GCCTGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTTCAGGCG 168 Lys_TTT_chr11: 59323902-59323974 (+) GCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGTC CGGGGTTCAAGTCCCTGTTCGGGCG 169 Lys_TTT_chr6: 27302769-27302841 (−) GCCTGGGTAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTCCAGGCG 170 Lys_TTT_chr6: 28715521-28715593 (+) GCCTGGATAGCTCAGTTGGTAGAACATCAGACTTTTAATCTGACGGTG CAGGGTTCAAGTCCCTGTTCAGGCG 171 Met_CAT_chr8: 124169470-124169542 (−) GCCTCGTTAGCGCAGTAGGTAGCGCGTCAGTCTCATAATCTGAAGGTC GTGAGTTCGATCCTCACACGGGGCA 172 Met_CAT_chr16: 71460396-71460468 (+) GCCCTCTTAGCGCAGTGGGCAGCGCGTCAGTCTCATAATCTGAAGGTC CTGAGTTCGAGCCTCAGAGAGGGCA 173 Met_CAT_chr6: 28912352-28912424 (+) GCCTCCTTAGCGCAGTAGGCAGCGCGTCAGTCTCATAATCTGAAGGTC CTGAGTTCGAACCTCAGAGGGGGCA 174 Met_CAT_chr6: 26735574-26735646 (−) GCCCTCTTAGCGCAGCGGGCAGCGCGTCAGTCTCATAATCTGAAGGTC CTGAGTTCGAGCCTCAGAGAGGGCA 175 Met_CAT_chr6: 26701712-26701784 (+) GCCCTCTTAGCGCAGCTGGCAGCGCGTCAGTCTCATAATCTGAAGGTC CTGAGTTCAAGCCTCAGAGAGGGCA 176 Met_CAT_chr16: 87417628-87417700 (−) GCCTCGTTAGCGCAGTAGGCAGCGCGTCAGTCTCATAATCTGAAGGTC GTGAGTTCGAGCCTCACACGGGGCA 177 Met_CAT_chr6: 58168492-58168564 (−) GCCCTCTTAGTGCAGCTGGCAGCGCGTCAGTTTCATAATCTGAAAGTCC TGAGTTCAAGCCTCAGAGAGGGCA 178 Phe_GAA_chr6: 28758499-28758571 (−) GCCGAAATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTAAAGGTC CCTGGTTCGATCCCGGGTTTCGGCA 179 Phe_GAA_chr11: 59333853-59333925 (−) GCCGAAATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTAAAGGTC CCTGGTTCAATCCCGGGTTTCGGCA 180 Phe_GAA_chr6: 28775610-28775682 (−) GCCGAGATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTAAAGGTC CCTGGTTCAATCCCGGGTTTCGGCA 181 Phe_GAA_chr6: 28791093-28791166 (−) GCCGAAATAGCTCAGTTGGGAGAGCGTTAGACCGAAGATCTTAAAGGT CCCTGGTTCAATCCCGGGTTTCGGCA 182 Phe_GAA_chr6: 28731374-28731447 (−) GCTGAAATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTTAAAGTT CCCTGGTTCAACCCTGGGTTTCAGCC 183 Pro_AGG_chr16: 3241989-3242060 (+) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTAGGATGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCC 184 Pro_AGG_chr1: 167684725-167684796 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTAGGGTGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCC 185 Pro_CGG_chr1: 167683962-167684033 (+) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTCGGGTGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCC 186 Pro_CGG_chr6: 27059521-27059592 (+) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTCGGGTGTGAGAGGTCCC GGGTTCAAATCCCGGACGAGCCC 187 Pro_TGG_chr14: 21101165-21101236 (+) GGCTCGTTGGTCTAGTGGTATGATTCTCGCTTTGGGTGCGAGAGGTCCC GGGTTCAAATCCCGGACGAGCCC 188 Pro_TGG_chr11: 75946869-75946940 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGGTTTGGGTCCGAGAGGTCCC GGGTTCAAATCCCGGACGAGCCC 189 Pro_TGG_chr5: 180615854-180615925 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTTGGGTGCGAGAGGTCCC GGGTTCAAATCCCGGACGAGCCC 190 SeC_TCA_chr19: 45981859-45981945 (−) GCCCGGATGATCCTCAGTGGTCTGGGGTGCAGGCTTCAAACCTGTAGC TGTCTAGCGACAGAGTGGTTCAATTCCACCTTTCGGGCG 191 SeC_TCA_chr22: 44546537-44546620 (+) GCTCGGATGATCCTCAGTGGTCTGGGGTGCAGGCTTCAAACCTGTAGC TGTCTAGTGACAGAGTGGTTCAATTCCACCTTTGTA 192 Ser_AGA_chr6: 27509554-27509635 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTAGAAATCCATTGGGG TTTCCCCGCGCAGGTTCGAATCCTGCCGACTACG 193 Ser_AGA_chr6: 26327817-26327898 (+) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTAGAAATCCATTGGGG TCTCCCCGCGCAGGTTCGAATCCTGCCGACTACG 194 Ser_AGA_chr6: 27499987-27500068 (+) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTAGAAATCCATTGGGG TTTCCCCACGCAGGTTCGAATCCTGCCGACTACG 195 Ser_AGA_chr6: 27521192-27521273 (−) GTAGTCGTGGCCGAGTGGTTAAGGTGATGGACTAGAAACCCATTGGGG TCTCCCCGCGCAGGTTCGAATCCTGCCGACTACG 196 Ser_CGA_chr17: 8042199-8042280 (−) GCTGTGATGGCCGAGTGGTTAAGGCGTTGGACTCGAAATCCAATGGGG TCTCCCCGCGCAGGTTCGAATCCTGCTCACAGCG 197 Ser_CGA_chr6: 27177628-27177709 (+) GCTGTGATGGCCGAGTGGTTAAGGCGTTGGACTCGAAATCCAATGGGG TCTCCCCGCGCAGGTTCAAATCCTGCTCACAGCG 198 Ser_CGA_chr6: 27640229-27640310 (−) GCTGTGATGGCCGAGTGGTTAAGGTGTTGGACTCGAAATCCAATGGGG GTTCCCCGCGCAGGTTCAAATCCTGCTCACAGCG 199 Ser_CGA_chr12: 56584148-56584229 (+) GTCACGGTGGCCGAGTGGTTAAGGCGTTGGACTCGAAATCCAATGGGG TTTCCCCGCACAGGTTCGAATCCTGTTCGTGACG 200 Ser_GCT_chr6: 27065085-27065166 (+) GACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTGC TCTGCACGCGTGGGTTCGAATCCCACCCTCGTCG 201 Ser_GCT_chr6: 27265775-27265856 (+) GACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTGC TCTGCACGCGTGGGTTCGAATCCCACCTTCGTCG 202 Ser_GCT_chr11: 66115591-66115672 (+) GACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTGC TTTGCACGCGTGGGTTCGAATCCCATCCTCGTCG 203 Ser_GCT_chr6: 28565117-28565198 (−) GACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTGC TCTGCACGCGTGGGTTCGAATCCCATCCTCGTCG 204 Ser_GCT_chr6: 28180815-28180896 (+) GACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTGC TCTGCACACGTGGGTTCGAATCCCATCCTCGTCG 205 Ser_GCT_chr6: 26305718-26305801 (−) GGAGAGGCCTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGT GCTCTGCACGCGTGGGTTCGAATCCCATCCTCGTCG 206 Ser_TGA_chr10: 69524261-69524342 (+) GCAGCGATGGCCGAGTGGTTAAGGCGTTGGACTTGAAATCCAATGGGG TCTCCCCGCGCAGGTTCGAACCCTGCTCGCTGCG 207 Ser_TGA_chr6: 27513468-27513549 (+) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTTGAAATCCATTGGGG TTTCCCCGCGCAGGTTCGAATCCTGCCGACTACG 208 Ser_TGA_chr6: 26312824-26312905 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTTGAAATCCATTGGGG TCTCCCCGCGCAGGTTCGAATCCTGCCGACTACG 209 Ser_TGA_chr6: 27473607-27473688 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTTGAAATCCATTGGGG TTTCCCCGCGCAGGTTCGAATCCTGTCGGCTACG 210 Thr_AGT_chr17: 8090478-8090551 (+) GGCGCCGTGGCTTAGTTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGTGCCT 211 Thr_AGT_chr6: 26533145-26533218 (−) GGCTCCGTGGCTTAGCTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGGGCCT 212 Thr_AGT_chr6: 28693795-28693868 (+) GGCTCCGTAGCTTAGTTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGACTCCCAGCGGGGCCT 213 Thr_AGT_chr6: 27694473-27694546 (+) GGCTTCGTGGCTTAGCTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGAGGCCT 214 Thr_AGT_chr17: 8042770-8042843 (−) GGCGCCGTGGCTTAGCTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGTGCCT 215 Thr_AGT_chr6: 27130050-27130123 (+) GGCCCTGTGGCTTAGCTGGTCAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGGGCCT 216 Thr_CGT_chr6: 28456770-28456843 (−) GGCTCTATGGCTTAGTTGGTTAAAGCGCCTGTCTCGTAAACAGGAGAT CCTGGGTTCGACTCCCAGTGGGGCCT 217 Thr_CGT_chr16: 14379750-14379821 (+) GGCGCGGTGGCCAAGTGGTAAGGCGTCGGTCTCGTAAACCGAAGATCA CGGGTTCGAACCCCGTCCGTGCCT 218 Thr_CGT_chr6: 28615984-28616057 (−) GGCTCTGTGGCTTAGTTGGCTAAAGCGCCTGTCTCGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGGGCCT 219 Thr_CGT_chr17: 29877093-29877164 (+) GGCGCGGTGGCCAAGTGGTAAGGCGTCGGTCTCGTAAACCGAAGATCG CGGGTTCGAACCCCGTCCGTGCCT 220 Thr_CGT_chr6: 27586135-27586208 (+) GGCCCTGTAGCTCAGCGGTTGGAGCGCTGGTCTCGTAAACCTAGGGGT CGTGAGTTCAAATCTCACCAGGGCCT 221 Thr_TGT_chr6: 28442329-28442402 (−) GGCTCTATGGCTTAGTTGGTTAAAGCGCCTGTCTTGTAAACAGGAGAT CCTGGGTTCGAATCCCAGTAGAGCCT 222 Thr_TGT_chr1: 222638347-222638419 (+) GGCTCCATAGCTCAGTGGTTAGAGCACTGGTCTTGTAAACCAGGGGTC GCGAGTTCGATCCTCGCTGGGGCCT 223 Thr_TGT_chr14: 21081949-21082021 (−) GGCTCCATAGCTCAGGGGTTAGAGCGCTGGTCTTGTAAACCAGGGGTC GCGAGTTCAATTCTCGCTGGGGCCT 224 Thr_TGT_chr14: 21099319-21099391 (−) GGCTCCATAGCTCAGGGGTTAGAGCACTGGTCTTGTAAACCAGGGGTC GCGAGTTCAAATCTCGCTGGGGCCT 225 Thr_TGT_chr14: 21149849-21149921 (+) GGCCCTATAGCTCAGGGGTTAGAGCACTGGTCTTGTAAACCAGGGGTC GCGAGTTCAAATCTCGCTGGGGCCT 226 Thr_TGT_chr5: 180618687-180618758 (−) GGCTCCATAGCTCAGGGGTTAGAGCACTGGTCTTGTAAACCAGGGTCG CGAGTTCAAATCTCGCTGGGGCCT 227 Trp_CCA_chr17: 8124187-8124258 (−) GGCCTCGTGGCGCAACGGTAGCGCGTCTGACTCCAGATCAGAAGGTTG CGTGTTCAAATCACGTCGGGGTCA 228 Trp_CCA_chr17: 19411494-19411565 (+) GACCTCGTGGCGCAATGGTAGCGCGTCTGACTCCAGATCAGAAGGTTG CGTGTTCAAGTCACGTCGGGGTCA 229 Trp_CCA_chr6: 26319330-26319401 (−) GACCTCGTGGCGCAACGGTAGCGCGTCTGACTCCAGATCAGAAGGTTG CGTGTTCAAATCACGTCGGGGTCA 230 Trp_CCA_chr12: 98898030-98898101 (+) GACCTCGTGGCGCAACGGTAGCGCGTCTGACTCCAGATCAGAAGGCTG CGTGTTCGAATCACGTCGGGGTCA 231 Trp_CCA_chr7: 99067307-99067378 (+) GACCTCGTGGCGCAACGGCAGCGCGTCTGACTCCAGATCAGAAGGTTG CGTGTTCAAATCACGTCGGGGTCA 232 Tyr_ATA_chr2: 219110549-219110641 (+) CCTTCAATAGTTCAGCTGGTAGAGCAGAGGACTATAGCTACTTCCTCA GTAGGAGACGTCCTTAGGTTGCTGGTTCGATTCCAGCTTGAAGGA 233 Tyr_GTA_chr6: 26569086-26569176 (+) CCTTCGATAGCTCAGTTGGTAGAGCGGAGGACTGTAGTTGGCTGTGTC CTTAGACATCCTTAGGTCGCTGGTTCGAATCCGGCTCGAAGGA 234 Tyr_GTA_chr2: 27273650-27273738 (+) CCTTCGATAGCTCAGTTGGTAGAGCGGAGGACTGTAGTGGATAGGGCG TGGCAATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 235 Tyr_GTA_chr6: 26577332-26577420 (+) CCTTCGATAGCTCAGTTGGTAGAGCGGAGGACTGTAGGCTCATTAAGC AAGGTATCCTTAGGTCGCTGGTTCGAATCCGGCTCGGAGGA 236 Tyr_GTA_chr14: 21125623-21125716 (−) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGATTGTATAGAC ATTTGCGGACATCCTTAGGTCGCTGGTTCGATTCCAGCTCGAAGGA 237 Tyr_GTA_chr8: 67025602-67025694 (+) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGCTACTTCCTCA GCAGGAGACATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 238 Tyr_GTA_chr8: 67026223-67026311 (+) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGGCGCGCGCCCG TGGCCATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 239 Tyr_GTA_chr14: 21121258-21121351 (−) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGCCTGTAGAAAC ATTTGTGGACATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 240 Tyr_GTA_chr14: 21131351-21131444 (−) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGATTGTACAGAC ATTTGCGGACATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 241 Tyr_GTA_chr14: 21151432-21151520 (+) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGTACTTAATGTG TGGTCATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 242 Tyr_GTA_chr6: 26595102-26595190 (+) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGGGGTTTGAATG TGGTCATCCTTAGGTCGCTGGTTCGAATCCGGCTCGGAGGA 243 Tyr_GTA_chr14: 21128117-21128210 (−) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGACTGCGGAAAC GTTTGTGGACATCCTTAGGTCGCTGGTTCAATTCCGGCTCGAAGGA 244 Tyr_GTA_chr6: 26575798-26575887 (+) CTTTCGATAGCTCAGTTGGTAGAGCGGAGGACTGTAGGTTCATTAAAC TAAGGCATCCTTAGGTCGCTGGTTCGAATCCGGCTCGAAGGA 245 Tyr_GTA_chr8: 66609532-66609619 (−) TCTTCAATAGCTCAGCTGGTAGAGCGGAGGACTGTAGGTGCACGCCCG TGGCCATTCTTAGGTGCTGGTTTGATTCCGACTTGGAGAG 246 Val_AAC_chr3: 169490018-169490090 (+) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGTCC CCGGTTCGAAACCGGGCGGAAACA 247 Val_AAC_chr5: 180615416-180615488 (−) GTTTCCGTAGTGTAGTGGTCATCACGTTCGCCTAACACGCGAAAGGTC CCCGGTTCGAAACCGGGCGGAAACA 248 Val_AAC_chr6: 27618707-27618779 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGTCC CTGGATCAAAACCAGGCGGAAACA 249 Val_AAC_chr6: 27648885-27648957 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGTCC GCGGTTCGAAACCGGGCGGAAACA 250 Val_AAC_chr6: 27203288-27203360 (+) GTTTCCGTAGTGTAGTGGTTATCACGTTTGCCTAACACGCGAAAGGTCC CCGGTTCGAAACCGGGCAGAAACA 251 Val_AAC_chr6: 28703206-28703277 (−) GGGGGTGTAGCTCAGTGGTAGAGCGTATGCTTAACATTCATGAGGCTC TGGGTTCGATCCCCAGCACTTCCA 252 Val_CAC_chr1: 161369490-161369562 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACCGGGCGGAAACA 253 Val_CAC_chr6: 27248049-27248121 (−) GCTTCTGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACCGGGCAGAAGCA 254 Val_CAC_chr19: 4724647-4724719 (−) GTTTCCGTAGTGTAGCGGTTATCACATTCGCCTCACACGCGAAAGGTCC CCGGTTCGATCCCGGGCGGAAACA 255 Val_CAC_chr1: 149298555-149298627 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACTGGGCGGAAACA 256 Val_CAC_chr1: 149684088-149684161 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGTAAAGGTC CCCGGTTCGAAACCGGGCGGAAACA 257 Val_CAC_chr6: 27173867-27173939 (−) GTTTCCGTAGTGGAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTC CCCGGTTTGAAACCAGGCGGAAACA 258 Val_TAC_chr11: 59318102-59318174 (−) GGTTCCATAGTGTAGTGGTTATCACGTCTGCTTTACACGCAGAAGGTCC TGGGTTCGAGCCCCAGTGGAACCA 259 Val_TAC_chr11: 59318460-59318532 (−) GGTTCCATAGTGTAGCGGTTATCACGTCTGCTTTACACGCAGAAGGTCC TGGGTTCGAGCCCCAGTGGAACCA 260 Val_TAC_chr10: 5895674-5895746 (−) GGTTCCATAGTGTAGTGGTTATCACATCTGCTTTACACGCAGAAGGTCC TGGGTTCAAGCCCCAGTGGAACCA 261 Val_TAC_chr6: 27258405-27258477 (+) GTTTCCGTGGTGTAGTGGTTATCACATTCGCCTTACACGCGAAAGGTCC TCGGGTCGAAACCGAGCGGAAACA 262 iMet_CAT_chr1: 153643726-153643797 (+) AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTC GATGGATCGAAACCATCCTCTGCTA 263 iMet_CAT_chr6: 27745664-27745735 (+) AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTC GATGGATCTAAACCATCCTCTGCTA 264 Glu_TTC_chr1: 16861773-16861845 (−) TCCCTGGTGGTCTAGTGGCTAGGATTCGGCGCTTTCACCGCCGCGGCCC GGGTTCGATTCCCGGTCAGGGAAT 265 Gly_CCC_chr1: 17004765-17004836 (−) GCGTTGGTGGTTTAGTGGTAGAATTCTCGCCTCCCATGCGGGAGACCC GGGTTCAATTCCCGGCCACTGCAC 266 Gly_CCC_chr1: 17053779-17053850 (+) GGCCTTGGTGGTGCAGTGGTAGAATTCTCGCCTCCCACGTGGGAGACC CGGGTTCAATTCCCGGCCAATGCA 267 Glu_TTC_chr1: 17199077-17199149 (+) GTCCCTGGTGGTCTAGTGGCTAGGATTCGGCGCTTTCACCGCCGCGGCC CGGGTTCGATTCCCGGCCAGGGAA 268 Asn_GTT_chr1: 17216171-17216245 (+) TGTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGA TTGGTGGTTCGAGCCCACCCAGGGACG 269 Arg_TCT_chr1: 94313128-94313213 (+) TGGCTCCGTGGCGCAATGGATAGCGCATTGGACTTCTAGAGGCTGAAG GCATTCAAAGGTTCCGGGTTCGAGTCCCGGCGGAGTCG 270 Lys_CTT_chr1: 145395521-145395594 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCGC 271 His_GTG_chr1: 145396880-145396952 (−) GCCGTGATCGTATAGTGGTTAGTACTCTGCGTTGTGGCCGCAGCAACCT CGGTTCGAATCCGAGTCACGGCAG 272 Gly_TCC_chr1: 145397863-145397935 (−) GCGTTGGTGGTATAGTGGTGAGCATAGCTGCCTTCCAAGCAGTTGACC CGGGTTCGATTCCCGGCCAACGCAG 273 Glu_CTC_chr1: 145399232-145399304 (−) TCCCTGGTGGTCTAGTGGTTAGGATTCGGCGCTCTCACCGCCGCGGCCC GGGTTCGATTCCCGGTCAGGGAAA 274 Gln_CTG_chr1: 145963303-145963375 (+) AGGTTCCATGGTGTAATGGTGAGCACTCTGGACTCTGAATCCAGCGAT CCGAGTTCGAGTCTCGGTGGAACCT 275 Asn_GTT_chr1: 148000804-148000878 (+) TGTCTCTGTGGCGTAGTCGGTTAGCGCGTTCGGCTGTTAACCGAAAAGT TGGTGGTTCGAGCCCACCCAGGAACG 276 Asn_GTT_chr1: 148248114-148248188 (+) TGTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGG TTGGTGGTTCGAGCCCACCCAGGGACG 277 Asn_GTT_chr1: 148598313-148598387 (−) GTCTCTGTGGCGCAATCGGTTAGCGCATTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCACCCAGGGACGC 278 Asn_GTT_chr1: 149230569-149230643 (−) GTCTCTGTGGCGCAATGGGTTAGCGCGTTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCATCCAGGGACGC 279 Val_CAC_chr1: 149294665-149294736 (−) GCACTGGTGGTTCAGTGGTAGAATTCTCGCCTCACACGCGGGACACCC GGGTTCAATTCCCGGTCAAGGCAA 280 Val_CAC_chr1: 149298554-149298627 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACTGGGCGGAAACAG 281 Gly_CCC_chr1: 149680209-149680280 (−) GCACTGGTGGTTCAGTGGTAGAATTCTCGCCTCCCACGCGGGAGACCC GGGTTTAATTCCCGGTCAAGATAA 282 Val_CAC_chr1: 149684087-149684161 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGTAAAGGTC CCCGGTTCGAAACCGGGCGGAAACAT 283 Met_CAT_chr1: 153643725-153643797 (+) TAGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGT CGATGGATCGAAACCATCCTCTGCTA 284 Val_CAC_chr1: 161369489-161369562 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACCGGGCGGAAACAA 285 Asp_GTC_chr1: 161410614-161410686 (−) TCCTCGTTAGTATAGTGGTGAGTATCCCCGCCTGTCACGCGGGAGACC GGGGTTCGATTCCCCGACGGGGAGG 286 Gly_GCC_chr1: 161413093-161413164 (+) TGCATGGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCC CGGGTTCGATTCCCGGCCCATGCA 287 Glu_CTC_chr1: 161417017-161417089 (−) TCCCTGGTGGTCTAGTGGTTAGGATTCGGCGCTCTCACCGCCGCGGCCC GGGTTCGATTCCCGGTCAGGGAAG 288 Asp_GTC_chr1: 161492934-161493006 (+) ATCCTTGTTACTATAGTGGTGAGTATCTCTGCCTGTCATGCGTGAGAGA GGGGGTCGATTCCCCGACGGGGAG 289 Gly_GCC_chr1: 161493636-161493707 (−) GCATTGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCCC GGGTTCGATTCCCGGCCAATGCAC 290 Leu_CAG_chr1: 161500131-161500214 (−) GTCAGGATGGCCGAGCGGTCTAAGGCGCTGCGTTCAGGTCGCAGTCTC CCCTGGAGGCGTGGGTTCGAATCCCACTCCTGACAA 291 Gly_TCC_chr1: 161500902-161500974 (+) CGCGTTGGTGGTATAGTGGTGAGCATAGCTGCCTTCCAAGCAGTTGAC CCGGGTTCGATTCCCGGCCAACGCA 292 Asn_GTT_chr1: 161510030-161510104 (+) CGTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGG TTGGTGGTTCGATCCCACCCAGGGACG 293 Glu_TTC_chr1: 161582507-161582579 (+) CGCGTTGGTGGTGTAGTGGTGAGCACAGCTGCCTTTCAAGCAGTTAAC GCGGGTTCGATTCCCGGGTAACGAA 294 Pro_CGG_chr1: 167683961-167684033 (+) CGGCTCGTTGGTCTAGGGGTATGATTCTCGCTTCGGGTGCGAGAGGTC CCGGGTTCAAATCCCGGACGAGCCC 295 Pro_AGG_chr1: 167684724-167684796 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTAGGGTGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCCT 296 Lys_TTT_chr1: 204475654-204475727 (+) CGCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGT CCAGGGTTCAAGTCCCTGTTCGGGCG 297 Lys_TTT_chr1: 204476157-204476230 (−) GCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTTCGGGCGT 298 Leu_CAA_chr1: 249168053-249168159 (+) TGTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGGTAAGCACC TTGCCTGCGGGCTTTCTGGTCTCCGGATGGAGGCGTGGGTTCGAATCCC 299 Glu_CTC_chr1: 249168446-249168518 (+) TTCCCTGGTGGTCTAGTGGTTAGGATTCGGCGCTCTCACCGCCGCGGCC CGGGTTCGATTCCCGGTCAGGAAA 300 Tyr_GTA_chr2: 27273649-27273738 (+) GCCTTCGATAGCTCAGTTGGTAGAGCGGAGGACTGTAGTGGATAGGGC GTGGCAATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 301 Ala_AGC_chr2: 27274081-27274154 (+) CGGGGGATTAGCTCAAATGGTAGAGCGCTCGCTTAGCATGCGAGAGGT AGCGGGATCGATGCCCGCATCCTCCA 302 Ile_TAT_chr2: 43037675-43037768 (+) AGCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATACAGCAGTAC ATGCAGAGCAATGCCGAGGTTGTGAGTTCGAGCCTCACCTGGAGCA 303 Gly_CCC_chr2: 70476122-70476193 (−) GCGCCGCTGGTGTAGTGGTATCATGCAAGATTCCCATTCTTGCGACCCG GGTTCGATTCCCGGGCGGCGCAT 304 Glu_TTC_chr2: 131094700-131094772 (−) TCCCATATGGTCTAGCGGTTAGGATTCCTGGTTTTCACCCAGGTGGCCC GGGTTCGACTCCCGGTATGGGAAC 305 Ala_CGC_chr2: 157257280-157257352 (+) GGGGGATGTAGCTCAGTGGTAGAGCGCGCGCTTCGCATGTGTGAGGTC CCGGGTTCAATCCCCGGCATCTCCA 306 Gly_GCC_chr2: 157257658-157257729 (−) GCATTGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCCC GGGTTCGATTCCCGGCCAATGCAA 307 Arg_ACG_chr3: 45730490-45730563 (−) GGGCCAGTGGCGCAATGGATAACGCGTCTGACTACGGATCAGAAGATT CTAGGTTCGACTCCTGGCTGGCTCGC 308 Val_AAC_chr3: 169490017-169490090 (+) GGTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGT CCCCGGTTCGAAACCGGGCGGAAACA 309 Val_AAC_chr5: 180596609-180596682 (+) AGTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGT CCCCGGTTCGAAACCGGGCGGAAACA 310 Leu_AAG_chr5: 180614700-180614782 (+) AGGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTAAGGCTCCAGTCT CTTCGGGGGCGTGGGTTCGAATCCCACCGCTGCCA 311 Val_AAC_chr5: 180615415-180615488 (−) GTTTCCGTAGTGTAGTGGTCATCACGTTCGCCTAACACGCGAAAGGTC CCCGGTTCGAAACCGGGCGGAAACAT 312 Pro_TGG_chr5: 180615853-180615925 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTTGGGTGCGAGAGGTCCC GGGTTCAAATCCCGGACGAGCCCA 313 Thr_TGT_chr5: 180618686-180618758 (−) GGCTCCATAGCTCAGGGGTTAGAGCACTGGTCTTGTAAACCAGGGTCG CGAGTTCAAATCTCGCTGGGGCCTG 314 Ala_TGC_chr5: 180633867-180633939 (+) TGGGGATGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGCC CCGGGTTCGATCCCCGGCATCTCCA 315 Lys_CTT_chr5: 180634754-180634827 (+) CGCCCGGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGT CGTGGGTTCGAGCCCCACGTTGGGCG 316 Val_AAC_chr5: 180645269-180645342 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGTCC CCGGTTCGAAACCGGGCGGAAACAA 317 Lys_CTT_chr5: 180648978-180649051 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCGT 318 Val_CAC_chr5: 180649394-180649467 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACCGGGCGGAAACAC 319 Met_CAT_chr6: 26286753-26286825 (+) CAGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGT CGATGGATCGAAACCATCCTCTGCTA 320 Ser_GCT_chr6: 26305717-26305801 (−) GGAGAGGCCTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGT GCTCTGCACGCGTGGGTTCGAATCCCATCCTCGTCGC 321 Gln_TTG_chr6: 26311423-26311495 (−) GGCCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCGATC CGAGTTCAAATCTCGGTGGGACCTG 322 Gln_TTG_chr6: 26311974-26312046 (−) GGCCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCGATC CGAGTTCAAATCTCGGTGGGACCTA 323 Ser_TGA_chr6: 26312823-26312905 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTTGAAATCCATTGGGG TCTCCCCGCGCAGGTTCGAATCCTGCCGACTACGG 324 Met_CAT_chr6: 26313351-26313423 (−) AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTC GATGGATCGAAACCATCCTCTGCTAT 325 Arg_TCG_chr6: 26323045-26323118 (+) GGACCACGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGAT TGAGGGTTCGAATCCCTCCGTGGTTA 326 Ser_AGA_chr6: 26327816-26327898 (+) TGTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTAGAAATCCATTGGG GTCTCCCCGCGCAGGTTCGAATCCTGCCGACTACG 327 Met_CAT_chr6: 26330528-26330600 (−) AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTC GATGGATCGAAACCATCCTCTGCTAG 328 Leu_CAG_chr6: 26521435-26521518 (+) CGTCAGGATGGCCGAGCGGTCTAAGGCGCTGCGTTCAGGTCGCAGTCT CCCCTGGAGGCGTGGGTTCGAATCCCACTCCTGACA 329 Thr_AGT_chr6: 26533144-26533218 (−) GGCTCCGTGGCTTAGCTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGGGCCTG 330 Arg_ACG_chr6: 26537725-26537798 (+) AGGGCCAGTGGCGCAATGGATAACGCGTCTGACTACGGATCAGAAGA TTCCAGGTTCGACTCCTGGCTGGCTCG 331 Val_CAC_chr6: 26538281-26538354 (+) GGTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTC CCCGGTTCGAAACCGGGCGGAAACA 332 Ala_CGC_chr6: 26553730-26553802 (+) AGGGGATGTAGCTCAGTGGTAGAGCGCATGCTTCGCATGTATGAGGTC CCGGGTTCGATCCCCGGCATCTCCA 333 Ile_AAT_chr6: 26554349-26554423 (+) TGGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGG TCGCGGGTTCGATCCCCGTACGGGCCA 334 Pro_AGG_chr6: 26555497-26555569 (+) CGGCTCGTTGGTCTAGGGGTATGATTCTCGCTTAGGGTGCGAGAGGTC CCGGGTTCAAATCCCGGACGAGCCC 335 Lys_CTT_chr6: 26556773-26556846 (+) AGCCCGGCTAGCTCAGTCGGTAGAGCATGAGACTCTTAATCTCAGGGT CGTGGGTTCGAGCCCCACGTTGGGCG 336 Tyr_GTA_chr6: 26569085-26569176 (+) TCCTTCGATAGCTCAGTTGGTAGAGCGGAGGACTGTAGTTGGCTGTGT CCTTAGACATCCTTAGGTCGCTGGTTCGAATCCGGCTCGAAGGA 337 Ala_AGC_chr6: 26572091-26572164 (−) GGGGAATTAGCTCAAATGGTAGAGCGCTCGCTTAGCATGCGAGAGGTA GCGGGATCGATGCCCGCATTCTCCAG 338 Met_CAT_chr6: 26766443-26766516 (+) CGCCCTCTTAGCGCAGCGGGCAGCGCGTCAGTCTCATAATCTGAAGGT CCTGAGTTCGAGCCTCAGAGAGGGCA 339 Ile_TAT_chr6: 26988124-26988218 (+) TGCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATATGGCAGTAT GTGTGCGAGTGATGCCGAGGTTGTGAGTTCGAGCCTCACCTGGAGCA 340 His_GTG_chr6: 27125905-27125977 (+) TGCCGTGATCGTATAGTGGTTAGTACTCTGCGTTGTGGCCGCAGCAACC TCGGTTCGAATCCGAGTCACGGCA 341 Ile_AAT_chr6: 27144993-27145067 (−) GGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGGT CGCGGGTTCGATCCCCGTACGGGCCAC 342 Val_AAC_chr6: 27203287-27203360 (+) AGTTTCCGTAGTGTAGTGGTTATCACGTTTGCCTAACACGCGAAAGGTC CCCGGTTCGAAACCGGGCAGAAACA 343 Val_CAC_chr6: 27248048-27248121 (−) GCTTCTGTAGTGTAGTGGTTATCACGTTCGCCTCACACGCGAAAGGTCC CCGGTTCGAAACCGGGCAGAAGCAA 344 Asp_GTC_chr6: 27447452-27447524 (+) TTCCTCGTTAGTATAGTGGTGAGTATCCCCGCCTGTCACGCGGGAGACC GGGGTTCGATTCCCCGACGGGGAG 345 Ser_TGA_chr6: 27473606-27473688 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTTGAAATCCATTGGGG TTTCCCCGCGCAGGTTCGAATCCTGTCGGCTACGG 346 Gln_CTG_chr6: 27487307-27487379 (+) AGGTTCCATGGTGTAATGGTTAGCACTCTGGACTCTGAATCCAGCGAT CCGAGTTCAAATCTCGGTGGAACCT 347 Asp_GTC_chr6: 27551235-27551307 (−) TCCTCGTTAGTATAGTGGTGAGTGTCCCCGTCTGTCACGCGGGAGACC GGGGTTCGATTCCCCGACGGGGAGA 348 Val_AAC_chr6: 27618706-27618779 (−) GTTTCCGTAGTGTAGTGGTTATCACGTTCGCCTAACACGCGAAAGGTCC CTGGATCAAAACCAGGCGGAAACAA 349 Ile_AAT_chr6: 27655966-27656040 (+) CGGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGG TCGCGGGTTCGATCCCCGTACTGGCCA 350 Gln_CTG_chr6: 27759134-27759206 (−) GGCCCCATGGTGTAATGGTCAGCACTCTGGACTCTGAATCCAGCGATC CGAGTTCAAATCTCGGTGGGACCCA 351 Gln_TTG_chr6: 27763639-27763711 (−) GGCCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCGATC CGAGTTCAAATCTCGGTGGGACCTT 352 Ala_AGC_chr6: 28574932-28575004 (+) TGGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGTACGAGGTC CCGGGTTCAATCCCCGGCACCTCCA 353 Ala_AGC_chr6: 28626013-28626085 (−) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTAGCATGCATGAGGTCC CGGGTTCGATCCCCAGCATCTCCAG 354 Ala_CGC_chr6: 28697091-28697163 (+) AGGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTCGCATGTACGAGGCC CCGGGTTCGACCCCCGGCTCCTCCA 355 Ala_AGC_chr6: 28806220-28806292 (−) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGCACGAGGCCC CGGGTTCAATCCCCGGCACCTCCAT 356 Ala_AGC_chr6: 28831461-28831533 (−) GGGGGTGTAGCTCAGTGGTAGAGCGCGTGCTTAGCATGCACGAGGCCC CGGGTTCAATCCCCGGCACCTCCAG 357 Leu_CAA_chr6: 28863999-28864105 (−) GTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGCTAAGCTTCC TCCGCGGTGGGGATTCTGGTCTCCAATGGAGGCGTGGGTTCGAATCCC 358 Leu_CAA_chr6: 28908829-28908934 (+) TGTCAGGATGGCCGAGTGGTCTAAGGCGCCAGACTCAAGCTTGGCTTC CTCGTGTTGAGGATTCTGGTCTCCAATGGAGGCGTGGGTTCGAATCCC 359 Gln_CTG_chr6: 28909377-28909449 (−) GGTTCCATGGTGTAATGGTTAGCACTCTGGACTCTGAATCCAGCGATCC GAGTTCAAATCTCGGTGGAACCTT 360 Leu_AAG_chr6: 28911398-28911480 (−) GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTAAGGCTCCAGTCTC TTCGGGGGCGTGGGTTCGAATCCCACCGCTGCCAG 361 Met_CAT_chr6: 28912351-28912424 (+) TGCCTCCTTAGCGCAGTAGGCAGCGCGTCAGTCTCATAATCTGAAGGT CCTGAGTTCGAACCTCAGAGGGGGCA 362 Lys_TTT_chr6: 28918805-28918878 (+) AGCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGT CCAGGGTTCAAGTCCCTGTTCGGGCG 363 Met_CAT_chr6: 28921041-28921114 (−) GCCTCCTTAGCGCAGTAGGCAGCGCGTCAGTCTCATAATCTGAAGGTC CTGAGTTCGAACCTCAGAGGGGGCAG 364 Glu_CTC_chr6: 28949975-28950047 (+) TTCCCTGGTGGTCTAGTGGTTAGGATTCGGCGCTCTCACCGCCGCGGCC CGGGTTCGATTCCCGGTCAGGGAA 365 Leu_TAA_chr6: 144537683-144537766 (+) CACCAGGATGGCCGAGTGGTTAAGGCGTTGGACTTAAGATCCAATGGA CATATGTCCGCGTGGGTTCGAACCCCACTCCTGGTA 366 Pro_AGG_chr7: 128423503-128423575 (+) TGGCTCGTTGGTCTAGGGGTATGATTCTCGCTTAGGGTGCGAGAGGTC CCGGGTTCAAATCCCGGACGAGCCC 367 Arg_CCT_chr7: 139025445-139025518 (+) AGCCCCAGTGGCCTAATGGATAAGGCATTGGCCTCCTAAGCCAGGGAT TGTGGGTTCGAGTCCCATCTGGGGTG 368 Cys_GCA_chr7: 149388271-149388343 (−) GGGGATATAGCTCAGGGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGGTGCCCCCCC 369 Tyr_GTA_chr8: 67025601-67025694 (+) CCCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGCTACTTCCTC AGCAGGAGACATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 370 Tyr_GTA_chr8: 67026222-67026311 (+) CCCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGGCGCGCGCCC GTGGCCATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 371 Ala_AGC_chr8: 67026423-67026496 (+) TGGGGGATTAGCTCAAATGGTAGAGCGCTCGCTTAGCATGCGAGAGGT AGCGGGATCGATGCCCGCATCCTCCA 372 Ser_AGA_chr8: 96281884-96281966 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTAGAAATCCATTGGGG TCTCCCCGCGCAGGTTCGAATCCTGCCGACTACGG 373 Met_CAT_chr8: 124169469-124169542 (−) GCCTCGTTAGCGCAGTAGGTAGCGCGTCAGTCTCATAATCTGAAGGTC GTGAGTTCGATCCTCACACGGGGCAC 374 Arg_TCT_chr9: 131102354-131102445 (−) GGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGCTGAGCCTAG TGTGGTCATTCAAAGGTTGTGGGTTCGAGTCCCACCAGAGTCGA 375 Asn_GTT_chr10: 22518437-22518511 (−) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCACCCAGGGACGC 376 Ser_TGA_chr10: 69524260-69524342 (+) GGCAGCGATGGCCGAGTGGTTAAGGCGTTGGACTTGAAATCCAATGGG GTCTCCCCGCGCAGGTTCGAACCCTGCTCGCTGCG 377 Val_TAC_chr11: 59318101-59318174 (−) GGTTCCATAGTGTAGTGGTTATCACGTCTGCTTTACACGCAGAAGGTCC TGGGTTCGAGCCCCAGTGGAACCAT 378 Val_TAC_chr11: 59318459-59318532 (−) GGTTCCATAGTGTAGCGGTTATCACGTCTGCTTTACACGCAGAAGGTCC TGGGTTCGAGCCCCAGTGGAACCAC 379 Arg_TCT_chr11: 59318766-59318852 (+) TGGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGATAGTTAGA GAAATTCAAAGGTTGTGGGTTCGAGTCCCACCAGAGTCG 380 Leu_TAA_chr11: 59319227-59319310 (+) TACCAGAATGGCCGAGTGGTTAAGGCGTTGGACTTAAGATCCAATGGA TTCATATCCGCGTGGGTTCGAACCCCACTTCTGGTA 381 Lys_TTT_chr11: 59323901-59323974 (+) GGCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGT CCGGGGTTCAAGTCCCTGTTCGGGCG 382 Phe_GAA_chr11: 59324969-59325042 (−) GCCGAAATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTAAAGGTC CCTGGTTCGATCCCGGGTTTCGGCAG 383 Lys_TTT_chr11:59327807-59327880 (−) GCCCGGATAGCTCAGTCGGTAGAGCATCAGACTTTTAATCTGAGGGTC CAGGGTTCAAGTCCCTGTTCGGGCGG 384 Phe_GAA_chr11:59333852-59333925 (−) GCCGAAATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTAAAGGTC CCTGGTTCAATCCCGGGTTTCGGCAG 385 Ser_GCT_chr11: 66115590-66115672 (+) GGACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTG CTTTGCACGCGTGGGTTCGAATCCCATCCTCGTCG 386 Pro_TGG_chr11: 75946868-75946940 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGGTTTGGGTCCGAGAGGTCCC GGGTTCAAATCCCGGACGAGCCCC 387 Ser_CGA_chr12: 56584147-56584229 (+) AGTCACGGTGGCCGAGTGGTTAAGGCGTTGGACTCGAAATCCAATGGG GTTTCCCCGCACAGGTTCGAATCCTGTTCGTGACG 388 Asp_GTC_chr12: 98897280-98897352 (+) CTCCTCGTTAGTATAGTGGTTAGTATCCCCGCCTGTCACGCGGGAGACC GGGGTTCAATTCCCCGACGGGGAG 389 Trp_CCA_chr12: 98898029-98898101 (+) GGACCTCGTGGCGCAACGGTAGCGCGTCTGACTCCAGATCAGAAGGCT GCGTGTTCGAATCACGTCGGGGTCA 390 Ala_TGC_chr12: 125406300-125406372 (−) GGGGATGTAGCTCAGTGGTAGAGCGCATGCTTTGCATGTATGAGGCCC CGGGTTCGATCCCCGGCATCTCCAT 391 Phe_GAA_chr12: 125412388-125412461 (−) GCCGAAATAGCTCAGTTGGGAGAGCGTTAGACTGAAGATCTAAAGGTC CCTGGTTCGATCCCGGGTTTCGGCAC 392 Ala_TGC_chr12: 125424511-125424583 (+) AGGGGATGTAGCTCAGTGGTAGAGCGCATGCTTTGCACGTATGAGGCC CCGGGTTCAATCCCCGGCATCTCCA 393 Asn_GTT_chr13: 31248100-31248174 (−) GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGGT TGGTGGTTCGAGCCCACCCAGGGACGG 394 Glu_TTC_chr13: 45492061-45492133 (−) TCCCACATGGTCTAGCGGTTAGGATTCCTGGTTTTCACCCAGGCGGCCC GGGTTCGACTCCCGGTGTGGGAAC 395 Thr_TGT_chr14: 21081948-21082021 (−) GGCTCCATAGCTCAGGGGTTAGAGCGCTGGTCTTGTAAACCAGGGGTC GCGAGTTCAATTCTCGCTGGGGCCTG 396 Leu_TAG_chr14: 21093528-21093610 (+) TGGTAGTGTGGCCGAGCGGTCTAAGGCGCTGGATTTAGGCTCCAGTCT CTTCGGGGGCGTGGGTTCGAATCCCACCACTGCCA 397 Thr_TGT_chr14: 21099318-21099391 (−) GGCTCCATAGCTCAGGGGTTAGAGCACTGGTCTTGTAAACCAGGGGTC GCGAGTTCAAATCTCGCTGGGGCCTC 398 Pro_TGG_chr14: 21101164-21101236 (+) TGGCTCGTTGGTCTAGTGGTATGATTCTCGCTTTGGGTGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCC 399 Tyr_GTA_chr14: 21131350-21131444 (−) CCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGATTGTACAGAC ATTTGCGGACATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGAA 400 Thr_TGT_chr14: 21149848-21149921 (+) AGGCCCTATAGCTCAGGGGTTAGAGCACTGGTCTTGTAAACCAGGGGT CGCGAGTTCAAATCTCGCTGGGGCCT 401 Tyr_GTA_chr14: 21151431-21151520 (+) TCCTTCGATAGCTCAGCTGGTAGAGCGGAGGACTGTAGTACTTAATGT GTGGTCATCCTTAGGTCGCTGGTTCGATTCCGGCTCGAAGGA 402 Pro_TGG_chr14: 21152174-21152246 (+) TGGCTCGTTGGTCTAGGGGTATGATTCTCGCTTTGGGTGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCC 403 Lys_CTT_chr14: 58706612-58706685 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGGGACTCTTAATCCCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCGC 404 Ile_AAT_chr14: 102783428-102783502 (+) CGGCCGGTTAGCTCAGTTGGTTAGAGCGTGGTGCTAATAACGCCAAGG TCGCGGGTTCGATCCCCGTACGGGCCA 405 Glu_TTC_chr15: 26327380-26327452 (−) TCCCACATGGTCTAGCGGTTAGGATTCCTGGTTTTCACCCAGGCGGCCC GGGTTCGACTCCCGGTGTGGGAAT 406 Ser_GCT_chr15: 40886022-40886104 (−) GACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTGC TCTGCACGCGTGGGTTCGAATCCCATCCTCGTCGA 407 His_GTG_chr15: 45490803-45490875 (−) GCCGTGATCGTATAGTGGTTAGTACTCTGCGTTGTGGCCGCAGCAACCT CGGTTCGAATCCGAGTCACGGCAT 408 His_GTG_chr15: 45493348-45493420 (+) CGCCGTGATCGTATAGTGGTTAGTACTCTGCGTTGTGGCCGCAGCAAC CTCGGTTCGAATCCGAGTCACGGCA 409 Gln_CTG_chr15: 66161399-66161471 (−) GGTTCCATGGTGTAATGGTTAGCACTCTGGACTCTGAATCCAGCGATCC GAGTTCAAATCTCGGTGGAACCTG 410 Lys_CTT_chr15: 79152903-79152976 (+) TGCCCGGCTAGCTCAGTCGGTAGAGCATGGGACTCTTAATCCCAGGGT CGTGGGTTCGAGCCCCACGTTGGGCG 411 Arg_TCG_chr15: 89878303-89878376 (+) GGGCCGCGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGAT TGCAGGTTCGAGTCCTGCCGCGGTCG 412 Gly_CCC_chr16: 686735-686806 (−) GCGCCGCTGGTGTAGTGGTATCATGCAAGATTCCCATTCTTGCGACCCG GGTTCGATTCCCGGGCGGCGCAC 413 Arg_CCG_chr16: 3200674-3200747 (+) GGGCCGCGTGGCCTAATGGATAAGGCGTCTGATTCCGGATCAGAAGAT TGAGGGTTCGAGTCCCTTCGTGGTCG 414 Arg_CCT_chr16: 3202900-3202973 (+) CGCCCCGGTGGCCTAATGGATAAGGCATTGGCCTCCTAAGCCAGGGAT TGTGGGTTCGAGTCCCACCCGGGGTA 415 Lys_CTT_chr16: 3207405-3207478 (−) GCCCGGCTAGCTCAGTCGGTAGAGCATGAGACCCTTAATCTCAGGGTC GTGGGTTCGAGCCCCACGTTGGGCGT 416 Thr_CGT_chr16: 14379749-14379821 (+) AGGCGCGGTGGCCAAGTGGTAAGGCGTCGGTCTCGTAAACCGAAGATC ACGGGTTCGAACCCCGTCCGTGCCT 417 Leu_TAG_chr16: 22207031-22207113 (−) GGTAGCGTGGCCGAGTGGTCTAAGGCGCTGGATTTAGGCTCCAGTCAT TTCGATGGCGTGGGTTCGAATCCCACCGCTGCCAC 418 Leu_AAG_chr16: 22308460-22308542 (+) GGGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTAAGGCTCCAGTCT CTTCGGGGGCGTGGGTTCGAATCCCACCGCTGCCA 419 Leu_CAG_chr16: 57333862-57333945 (+) AGTCAGGATGGCCGAGCGGTCTAAGGCGCTGCGTTCAGGTCGCAGTCT CCCCTGGAGGCGTGGGTTCGAATCCCACTTCTGACA 420 Leu_CAG_chr16: 57334391-57334474 (−) GTCAGGATGGCCGAGCGGTCTAAGGCGCTGCGTTCAGGTCGCAGTCTC CCCTGGAGGCGTGGGTTCGAATCCCACTTCTGACAG 421 Met_CAT_chr16: 87417627-87417700 (−) GCCTCGTTAGCGCAGTAGGCAGCGCGTCAGTCTCATAATCTGAAGGTC GTGAGTTCGAGCCTCACACGGGGCAG 422 Leu_TAG_chr17: 8023631-8023713 (−) GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGGATTTAGGCTCCAGTCTC TTCGGAGGCGTGGGTTCGAATCCCACCGCTGCCAG 423 Arg_TCT_chr17: 8024242-8024330 (+) TGGCTCTGTGGCGCAATGGATAGCGCATTGGACTTCTAGTGACGAATA GAGCAATTCAAAGGTTGTGGGTTCGAATCCCACCAGAGTCG 424 Gly_GCC_chr17: 8029063-8029134 (+) CGCATTGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCC CGGGTTCGATTCCCGGCCAATGCA 425 Ser_CGA_chr17: 8042198-8042280 (−) GCTGTGATGGCCGAGTGGTTAAGGCGTTGGACTCGAAATCCAATGGGG TCTCCCCGCGCAGGTTCGAATCCTGCTCACAGCGT 426 Thr_AGT_chr17: 8042769-8042843 (−) GGCGCCGTGGCTTAGCTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGTGCCTG 427 Trp_CCA_chr17: 8089675-8089747 (+) CGACCTCGTGGCGCAACGGTAGCGCGTCTGACTCCAGATCAGAAGGTT GCGTGTTCAAATCACGTCGGGGTCA 428 Ser_GCT_chr17: 8090183-8090265 (+) AGACGAGGTGGCCGAGTGGTTAAGGCGATGGACTGCTAATCCATTGTG CTCTGCACGCGTGGGTTCGAATCCCATCCTCGTCG 429 Thr_AGT_chr17: 8090477-8090551 (+) CGGCGCCGTGGCTTAGTTGGTTAAAGCGCCTGTCTAGTAAACAGGAGA TCCTGGGTTCGAATCCCAGCGGTGCCT 430 Trp_CCA_chr17: 8124186-8124258 (−) GGCCTCGTGGCGCAACGGTAGCGCGTCTGACTCCAGATCAGAAGGTTG CGTGTTCAAATCACGTCGGGGTCAA 431 Gly_TCC_chr17: 8124865-8124937 (+) AGCGTTGGTGGTATAGTGGTAAGCATAGCTGCCTTCCAAGCAGTTGAC CCGGGTTCGATTCCCGGCCAACGCA 432 Asp_GTC_chr17: 8125555-8125627 (−) TCCTCGTTAGTATAGTGGTGAGTATCCCCGCCTGTCACGCGGGAGACC GGGGTTCGATTCCCCGACGGGGAGA 433 Pro_CGG_chr17: 8126150-8126222 (−) GGCTCGTTGGTCTAGGGGTATGATTCTCGCTTCGGGTGCGAGAGGTCC CGGGTTCAAATCCCGGACGAGCCCT 434 Thr_AGT_chr17: 8129552-8129626 (−) GGCGCCGTGGCTTAGTTGGTTAAAGCGCCTGTCTAGTAAACAGGAGAT CCTGGGTTCGAATCCCAGCGGTGCCTT 435 Ser_AGA_chr17: 8129927-8130009 (−) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTAGAAATCCATTGGGG TCTCCCCGCGCAGGTTCGAATCCTGCCGACTACGT 436 Trp_CCA_chr17: 19411493-19411565 (+) TGACCTCGTGGCGCAATGGTAGCGCGTCTGACTCCAGATCAGAAGGTT GCGTGTTCAAGTCACGTCGGGGTCA 437 Thr_CGT_chr17: 29877092-29877164 (+) AGGCGCGGTGGCCAAGTGGTAAGGCGTCGGTCTCGTAAACCGAAGATC GCGGGTTCGAACCCCGTCCGTGCCT 438 Cys_GCA_chr17: 37023897-37023969 (+) AGGGGGTATAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTC CCCGGTTCAAATCCGGGTGCCCCCT 439 Cys_GCA_chr17: 37025544-37025616 (−) GGGGGTATAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CTGGTTCAAATCCGGGTGCCCCCTC 440 Cys_GCA_chr17: 37309986-37310058 (−) GGGGGTATAGCTCAGTGGTAGAGCATTTGACTGCAGATCAAGAGGTCC CCGGTTCAAATCCGGGTGCCCCCTC 441 Gln_TTG_chr17: 47269889-47269961 (+) AGGTCCCATGGTGTAATGGTTAGCACTCTGGACTTTGAATCCAGCGAT CCGAGTTCAAATCTCGGTGGGACCT 442 Arg_CCG_chr17: 66016012-66016085 (−) GACCCAGTGGCCTAATGGATAAGGCATCAGCCTCCGGAGCTGGGGATT GTGGGTTCGAGTCCCATCTGGGTCGC 443 Arg_CCT_chr17: 73030000-73030073 (+) AGCCCCAGTGGCCTAATGGATAAGGCACTGGCCTCCTAAGCCAGGGAT TGTGGGTTCGAGTCCCACCTGGGGTA 444 Arg_CCT_chr17: 73030525-73030598 (−) GCCCCAGTGGCCTAATGGATAAGGCACTGGCCTCCTAAGCCAGGGATT GTGGGTTCGAGTCCCACCTGGGGTGT 445 Arg_TCG_chr17: 73031207-73031280 (+) AGACCGCGTGGCCTAATGGATAAGGCGTCTGACTTCGGATCAGAAGAT TGAGGGTTCGAGTCCCTTCGTGGTCG 446 Asn_GTT_chr19: 1383561-1383635 (+) CGTCTCTGTGGCGCAATCGGTTAGCGCGTTCGGCTGTTAACCGAAAGG TTGGTGGTTCGAGCCCACCCAGGGACG 447 Gly_TCC_chr19: 4724081-4724153 (+) GGCGTTGGTGGTATAGTGGTTAGCATAGCTGCCTTCCAAGCAGTTGAC CCGGGTTCGATTCCCGGCCAACGCA 448 Val_CAC_chr19: 4724646-4724719 (−) GTTTCCGTAGTGTAGCGGTTATCACATTCGCCTCACACGCGAAAGGTCC CCGGTTCGATCCCGGGCGGAAACAG 449 Thr_AGT_chr19: 33667962-33668036 (+) TGGCGCCGTGGCTTAGTTGGTTAAAGCGCCTGTCTAGTAAACAGGAGA TCCTGGGTTCGAATCCCAGCGGTGCCT 450 Ile_TAT_chr19: 39902807-39902900 (−) GCTCCAGTGGCGCAATCGGTTAGCGCGCGGTACTTATATGACAGTGCG AGCGGAGCAATGCCGAGGTTGTGAGTTCGATCCTCACCTGGAGCAC 451 Gly_GCC_chr21: 18827106-18827177 (−) GCATGGGTGGTTCAGTGGTAGAATTCTCGCCTGCCACGCGGGAGGCCC GGGTTCGATTCCCGGCCCATGCAG

[0444] In an embodiment, a TREM, e.g., an exogenous TREM, comprises 1, 2, 3, or 4 of the following properties:

[0445] (a) differs by at least one nucleotide or one post transcriptional modification from the closest sequence tRNA in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced;

[0446] (b) has been introduced into a cell other than the cell in which it was transcribed;

[0447] (c) is present in a cell other than one in which it naturally occurs; or

[0448] (d) has an expression profile, e.g., level or distribution, that is non-wildtype, e.g., it is expressed at a higher level than wildtype.

[0449] In an embodiment, the expression profile can be mediated by a change introduced into a nucleic acid that modulates expression, or by addition of an agent that modulates expression of the RNA molecule.

[0450] In an embodiment, a TREM, e.g., an exogenous TREM comprises (a), (b), (c) and (d).

[0451] In an embodiment, a TREM, e.g., an exogenous TREM comprises (a), (b) and (c).

[0452] In an embodiment, a TREM, e.g., an exogenous TREM comprises (a), (b) and (d).

[0453] In an embodiment, a TREM, e.g., an exogenous TREM comprises (a), (c) and (d).

[0454] In an embodiment, a TREM, e.g., an exogenous TREM comprises (b), (c) and (d).

[0455] In an embodiment, a TREM, e.g., an exogenous TREM comprises (a) and (d).

[0456] In an embodiment, a TREM, e.g., an exogenous TREM comprises (c) and (d).

TREM Fragments

[0457] In an embodiment, a TREM comprises a fragment (sometimes referred to herein as a TREM fragment), e.g., a fragment of a RNA encoded by a deoxyribonucleic acid sequence disclosed in Table 1. E.g., the TREM includes less than the full sequence of a tRNA, e.g., less than the full sequence of a tRNA with the same anticodon, from the same species as the subject being treated, or both. In an embodiment, the production of a TREM fragment, e.g., from a full length TREM or a longer fragment, can be catalyzed by an enzyme, e.g., an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., Dicer, Angiogenin, RNaseP, RNaseZ, Rny1, or PrrC.

[0458] In an embodiment, a TREM fragment can be produced in vivo, ex vivo or in vitro. In an embodiment, a TREM fragment is produced in vivo, in the host cell. In an embodiment, a TREM fragment is produced ex vivo. In an embodiment, a TREM fragment is produced in vitro, e.g., as described in Example 12. In an embodiment, the TREM fragment is produced by fragmenting an expressed TREM after production of the TREM by the cell, e.g., a TREM produced by the host cell is fragmented after release or purification from the host cell, e.g., the TREM is fragmented ex vivo or in vitro.

[0459] Exemplary TREM fragments include TREM halves (e.g., from a cleavage in the ACHD, e.g., 5′TREM halves or 3′ TREM halves), a 5′ fragment (e.g., a fragment comprising the 5′ end, e.g., from a cleavage in a DHD or the ACHD), a 3′ fragment (e.g., a fragment comprising the 3′ end of a TREM, e.g., from a cleavage in the THD), or an internal fragment (e.g., from a cleavage in one or more of the ACHD, DHD or THD).

[0460] In an embodiment, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0461] In an embodiment, a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence with at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identity to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.

[0462] In an embodiment, a TREM fragment comprises a sequence of a length of between 10-90 ribonucleotides (rnt), between 10-80 rnt, between 10-70 rnt, between 10-60 rnt, between 10-50 rnt, between 10-40 rnt, between 10-30 rnt, between 10-20 rnt, between 20-90 rnt, between 20-80 rnt, 20-70 rnt, between 20-60 rnt, between 20-50 rnt, between 20-40 rnt, between 30-90 rnt, between 30-80 rnt, between 30-70 rnt, between 30-60 rnt, or between 30-50 rnt.

[0463] In an embodiment, a TREM fragment comprises a TREM structure, domain, or activity, e.g., as described herein above. In an embodiment, a TREM fragment comprises adaptor function, e.g., as described herein. In an embodiment, a TREM fragment comprises cognate adaptor function, e.g., as described herein. In an embodiment, a TREM fragment comprises non-cognate adaptor function, e.g., as described herein. In an embodiment, a TREM fragment comprises regulatory function, e.g., as described herein.

[0464] In an embodiment, a TREM fragment comprises translation inhibition function, e.g., displacement of an initiation factor, e.g., eIF4G.

[0465] In an embodiment, a TREM fragment comprises epigenetic function, e.g., epigenetic inheritance of a disorder, e.g., a metabolic disorder. In some embodiments, an epigenetic inheritance function can have a generational impact, e.g., as compared to somatic epigenetic regulation.

[0466] In an embodiment, a TREM fragment comprises retroviral regulation function, e.g., regulation of retroviral reverse transcription, e.g., HERV regulation.

[0467] In an embodiment, a TREM fragment comprises gene silencing function, e.g., by binding to AGO and/or PIWI.

[0468] In an embodiment, a TREM fragment comprises neuroprotectant function, e.g., by the sequestration of a translation initiation factor, e.g., in stress granules, to promote, e.g., motor neuron survival under cellular stress.

[0469] In an embodiment, a TREM fragment comprises anti-cancer function, e.g., by preventing cancer progression through the binding and/or sequestration of, e.g., metastatic transcript-stabilizing proteins.

[0470] In an embodiment, a TREM fragment comprises cell survival function, e.g., increased cell survival, by binding to, e.g., cytochrome c and/or cyt c ribonucleoprotein complex.

[0471] In an embodiment, a TREM fragment comprises ribosome biogenesis function, e.g., a TREM fragment can regulate ribosome biogenesis by, e.g., regulation of, e.g., binding to, an mRNA coding for ribosomal proteins.

TREM Modifications

[0472] A TREM described herein can comprise a moiety, often referred to herein as a modification, e.g., a moiety described in Table 2. While the term modification as used herein should not generally be construed to be the product of any particular process, in embodiments, the formation of a modification can be mediated by an enzyme in Table 2. In embodiments, the modification is formed post-transcriptionally. In embodiments, the modification is formed co-transcriptionally. In an embodiment, the modification occurs in vivo, e.g., in the host cell.

[0473] In an embodiment, the modification is a modification listed in any of rows 1-62 of Table 2. In an embodiment, the modification is a modification listed in any of rows 1-62 of Table 2, and the formation of the modification is mediated by an enzyme in Table 2. In an embodiment the modification is selected from a row in Table 2 and the formation of the modification is mediated by an enzyme from the same row in Table 2.

TABLE-US-00002 TABLE 2 List of tRNA modifications and associated enzymes. Short Name Modification Enzyme list 1 m1Am 1,2′-O-dimethyladenosine METTL3 2 imG wyosine Trm5, Tyw1, Tyw2, Tyw3, and Tyw4 3 m5s2U 5-methyl-2-thiouridine TrmU 4 m6t6A N6-methyl-N6- TRMO, TrmO threonylcarbamoyladenosine 5 QtRNA queuosine TGTase 6 OHyW hydroxywybutosine Trm5, TYW1 , TYW2, TYW3 , TYW4 7 io6A N6-(cis-hydroxyisopentenyl)adenosine TRIT1 8 Gr(p) 2′-O-ribosylguanosine (phosphate) 9 ho5U 5-hydroxyuridine 10 ncm5Um 5-carbamoylmethyl-2′-O-methyluridine ELP1, ELP2, ELP3, ELP4, ELP5, ELP6, KTI111, KTI112, KTI113, Uba4, Urm1, Tum1, Ncs6, Ncs2, Trm9, Sit4, Isu1, Isu2, Sap185, Sap190 11 OHyW* hydroxywybutosine wybutosine hydroxylases 12 acp3U 3-(3-amino-3-carboxypropyl)uridine 13 mcm5s2U 5-methoxycarbonylmethyl-2-thiouridine ALKBH8, Ncs6, Trm9, Ncs2, TrmU, CTU1, CTU2, ELP1, ELP2, ELP3, ELP4, ELP5, ELP6 14 m5U 5-methyluridine Trm2 15 D dihydrouridine DUS1, DUS2, DUS3, DUS4 16 mcm5Um 5-methoxycarbonylmethyl-2′-O- ELP1, ELP2, ELP3, ELP4, ELP5, methyluridine ELP6, Trm9, ALKBH-MT,? 17 m5C 5-methylcytidine Dnmt2, Dnmt2, EfmM, Nop2, Rcm1, RlmI, RlmO, RsmB, RsmF, Trm4, nsun2 18 ac4C N4-acetylcytidine NAT10, Rra1, TmcA 19 m1A 1-methyladenosine Bmt2, KamB, NpmA, Rrp8, TRMT10C, Trm61, TrmI, TrmK, Trmt61A, Trmt61B 20 tm5U 5-taurinomethyluridine MTU1 21 m1G 1-methylguanosine AviRa, RImA(I), RlmA(II), TRM5, TRMT10A, TRMT10B, TRMT10C, Taw22, Trm10, Trm5, Trmb, TrmD 22 Cm 2-O-methylcytidine 23 m1I 1-methylinosine 24 Ar(p) 2′O-ribosyladenosine (phosphate) 25 galQtRNA galactosyl-queuosine 26 mcm5U 5-methoxycarbonylmethyluridine ALKBH8, Trm9, ELP1, ELP2, ELP3, ELP4, ELP5, ELP6 27 m1Y 1-methylpseudouridine 28 Gm 2′O-methylguanosine MRM1, Mrm1, Nop1, RNMTL1, RlmB, Spb1, Trm3, Trm7, TrmH 29 manQtRNA mannosyl-queuosine Man/Gal-Q-transferase 30 yW wybutosine TYW1, 2, 3, 4 31 f5C 5-formylcytidine MTU1 32 tm5s2U 5-taurinomethyl-2-thiouridine TrmU 33 m2, 2G N2,N2-dimethylguanosine Trm1 34 chm5U 5-carboxyhydroxymethyluridine 35 s2U 2-thiouridine MnmA, Mtu1, Ncs2, Ncs6, TrmU 36 mnm5s2U 5-methylaminomethyl-2-thiouridine MnmCD, MnmD, MnmA, Mtu1, TrmU 37 m6A N6-methyladenosine ErmAM, ErmBC, ErmC′, Ime4, METTL14, METTL3, RlmF, RlmJ, RsmA, TrmM 38 mchm5U 5-(carboxyhydroxymethyl)uridine methyl ALKBH8 ester 39 m2G N2-methylguanosine Trm112, Trm11 40 cmnm5U 5-carboxymethylaminomethyluridine tRNA (cytidine(34)-2′-O)-methyltransferase 41 Ym 2′O-methylpseudouridine NEP1 42 f5Cm 5-formyl-2′-O-methylcytidine 43 ncm5U 5-carbamoylmethyluridine ELP1, ELP2, ELP3, ELP4, ELP5, ELP6 44 I inosine Tad1, Tad2, Tad3, TadA 45 g6A N6-glycinylcarbamoyladenosine METTL8 46 cmnm5s2U 5-carboxymethylaminomethyl-2- MnmA, Mtu1, TrmU, MnmE, MnmG, thiouridine Mss1, Mto1 47 Um 2′O-methyluridine AviRb, MRM2, Mrm2, Nop1, RlmE, Spb1, Trm44, TrmJ, TrmL, aTrm56 48 Y pseudouridine Cbf5, Pus1, Pus10, Pus2, Pus3, Pus4, Pus5, Pus6, Pus7, Pus8, Pus9, RluA, RluB, RluC, RluD, RluE, RluF, TruA, TruB, TruC, TruD 49 ms2i6A 2-methylthio-N6-isopentenyladenosine MiaA 50 m3C 3 -methylcytidine Trm140, METTL2 and METTE6 51 o2yW peroxywybutosine TRM5, TYW1, TYW2, TYW3, TYW4, TYW5, TRM4 52 m5Um 5,2′O-dimethyluridine 53 ms2t6A 2-methylthio-N6- Yrdc/Sua5, MtaB/e-MtaB, SAM, “S” threonylcarbamoyladenosine 54 i6A N6-isopentenyladenosine MiaA, Mod5 55 ms2io6A 2-methylthio-N6-(cis- MiaE hydroxyisopentenyl) adenosine 56 Am 2_-O-methyladenosine (2′-O-methyladenosine-N6-)- methyltransferase 57 m7G 7-methylguanosine Abd1, ArmA, Bud23, RlmKL, RmtB, RsmG, Sgm, TRMB, Trm8, TrmB, WDR4 58 t6A N6-threonylcarbamoyladenosine Bud32, Gon7, Cgi121 59 N1-methylguanine Trm10 60 N7-methylguanine Trm8, Trm82 61 2′-O methylribose Trm3, Trm13, Trm44, Trm7, Trm732, Rtt10 62 Ribose 2′-O-ribosyl phosphate Rit1

TREM Fusion

[0474] In an embodiment, a TREM disclosed herein comprises an additional moiety, e.g., a fusion moiety. In an embodiment, the fusion moiety can be used for purification, to alter folding of the TREM, or as a targeting moiety. In an embodiment, the fusion moiety can comprise a tag, a linker, can be cleavable or can include a binding site for an enzyme. In an embodiment, the fusion moiety can be disposed at the N terminal of the TREM or at the C terminal of the TREM. In an embodiment, the fusion moiety can be encoded by the same or different nucleic acid molecule that encodes the TREM.

TREM Consensus Sequence

[0475] In an embodiment, a TREM disclosed herein comprises a consensus sequence provided herein.

[0476] In an embodiment, a TREM disclosed herein comprises a consensus sequence of Formula I.sub.ZZZ, wherein .sub.ZZZ indicates any of the twenty amino acids and Formula I corresponds to all species.

[0477] In an embodiment, a TREM disclosed herein comprises a consensus sequence of Formula II.sub.ZZZ, wherein .sub.ZZZ indicates any of the twenty amino acids and Formula II corresponds to mammals.

[0478] In an embodiment, a TREM disclosed herein comprises a consensus sequence of Formula III.sub.ZZZ, wherein .sub.ZZZ indicates any of the twenty amino acids and Formula III corresponds to humans.

[0479] In an embodiment, .sub.ZZZ indicates any of the twenty amino acids: Alanine, Arginine, Asparagine, Aspartate, Cysteine, Glutamine, Glutamate, Glycine, Histidine, Isoleucine, Methionine, Leucine, Lysine, Phenylalanine, Proline, Serine, Threonine, Tryptophan, Tyrosine, or Valine.

[0480] In an embodiment, a TREM disclosed herein comprises a property selected from the following:

[0481] a) under physiological conditions residue R.sub.0 forms a linker region, e.g., a Linker 1 region;

[0482] b) under physiological conditions residues R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7 and residues R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71 form a stem region, e.g., an AStD stem region;

[0483] c) under physiological conditions residues R.sub.8-R.sub.9 forms a linker region, e.g., a Linker 2 region;

[0484] d) under physiological conditions residues -R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14 R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28 form a stem-loop region, e.g., a D arm Region; e) under physiological conditions residue -R.sub.29 forms a linker region, e.g., a Linker 3 Region;

[0485] f) under physiological conditions residues -R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46 form a stem-loop region, e.g., an AC arm region;

[0486] g) under physiological conditions residue -[R.sub.47].sub.x comprises a variable region, e.g., as described herein;

[0487] h) under physiological conditions residues -R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64 form a stem-loop region, e.g., a T arm Region; or

[0488] i) under physiological conditions residue R.sub.72 forms a linker region, e.g., a Linker 4 region.

Alanine TREM Consensus Sequence

[0489] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.ALA (SEQ ID NO: 562), [0490] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72
wherein R is a ribonucleotide residue and the consensus for Ala is: [0491] R.sub.0=absent; [0492] R.sub.14, R.sub.57=are independently A or absent; [0493] R.sub.26=A, C, G or absent; [0494] R.sub.5, R.sub.6, R.sub.15, R.sub.16, R.sub.21, R.sub.30, R.sub.31, R.sub.32, R.sub.34, R.sub.37, R.sub.41, R.sub.42, R.sub.43, R.sub.44, R.sub.45, R.sub.48, R.sub.49, R.sub.50, R.sub.58, R.sub.59, [0495] R.sub.63, R.sub.64, R.sub.66, R.sub.67=are independently N or absent; [0496] R.sub.11, R.sub.35, R.sub.65=are independently A, C, U or absent; [0497] R.sub.1, R.sub.9, R.sub.20, R.sub.38, R.sub.40, R.sub.51, R.sub.52, R.sub.56=are independently A, G or absent; [0498] R.sub.7, R.sub.22, R.sub.25, R.sub.27, R.sub.29, R.sub.46, R.sub.53, R.sub.72=are independently A, G, U or absent; [0499] R.sub.24, R.sub.69=are independently A, U or absent; [0500] R.sub.70, R.sub.71=are independently C or absent; [0501] R.sub.3, R.sub.4=are independently C, G or absent; [0502] R.sub.12, R.sub.33, R.sub.36, R.sub.62, R.sub.68=are independently C, G, U or absent; [0503] R.sub.13, R.sub.17, R.sub.28, R.sub.39, R.sub.55, R.sub.60, R.sub.61=are independently C, U or absent; [0504] R.sub.10, R.sub.19, R.sub.23=are independently G or absent; [0505] R.sub.2=G, U or absent; [0506] R.sub.8, R.sub.18, R.sub.54=are independently U or absent; [0507] [R.sub.47].sub.x=N or absent;

[0508] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0509] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.ALA (SEQ ID NO: 563), [0510] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72
wherein R is a ribonucleotide residue and the consensus for Ala is: [0511] R.sub.0, R.sub.18=are absent; [0512] R.sub.14, R.sub.24, R.sub.57=are independently A or absent; [0513] R.sub.15, R.sub.26, R.sub.64=are independently A, C, G or absent; [0514] R.sub.16, R.sub.31, R.sub.50, R.sub.59=are independently N or absent; [0515] R.sub.11, R.sub.32, R.sub.37, R.sub.41, R.sub.43, R.sub.45, R.sub.49, R.sub.65, R.sub.66=are independently A, C, U or absent; [0516] R.sub.1, R.sub.5, R.sub.9, R.sub.25, R.sub.27, R.sub.38, R.sub.40, R.sub.46, R.sub.51, R.sub.56=are independently A, G or absent; [0517] R.sub.7, R.sub.22, R.sub.29, R.sub.42, R.sub.44, R.sub.53, R.sub.63, R.sub.72=are independently A, G, U or absent; [0518] R.sub.6, R.sub.35, R.sub.69=are independently A, U or absent; [0519] R.sub.55, R.sub.60, R.sub.70, R.sub.71=are independently C or absent; [0520] R.sub.3=C, G or absent; [0521] R.sub.12, R.sub.36, R.sub.48=are independently C, G, U or absent; [0522] R.sub.13, R.sub.17, R.sub.28, R.sub.30, R.sub.34, R.sub.39, R.sub.58, R.sub.61, R.sub.62, R.sub.67, R.sub.68=are independently C, U or absent; [0523] R.sub.4, R.sub.10, R.sub.19, R.sub.20, R.sub.23, R.sub.52=are independently G or absent; [0524] R.sub.2, R.sub.8, R.sub.33=are independently G, U or absent; [0525] R.sub.21, R.sub.54=are independently U or absent; [0526] [R.sub.47].sub.x=N or absent;

[0527] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0528] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.ALA (SEQ ID NO: 564),

[0529] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ala is: [0530] R.sub.0, R.sub.18=are absent; [0531] R.sub.14, R.sub.24, R.sub.57, R.sub.72=are independently A or absent; [0532] R.sub.15, R.sub.26, R.sub.64=are independently A, C, G or absent; [0533] R.sub.16, R.sub.31, R.sub.50=are independently N or absent; [0534] R.sub.11, R.sub.32, R.sub.37, R.sub.41, R.sub.43, R.sub.45, R.sub.49, R.sub.65, R.sub.66=are independently A, C, U or absent; [0535] R.sub.5, R.sub.9, R.sub.25, R.sub.27, R.sub.38, R.sub.40, R.sub.46, R.sub.51, R.sub.56=are independently A, G or absent; [0536] R.sub.7, R.sub.22, R.sub.29, R.sub.42, R.sub.44, R.sub.53, R.sub.63=are independently A, G, U or absent; [0537] R.sub.6, R.sub.35=are independently A, U or absent; [0538] R.sub.55, R.sub.60, R.sub.61, R.sub.70, R.sub.71=are independently C or absent; [0539] R.sub.12, R.sub.48, R.sub.59=are independently C, G, U or absent; [0540] R.sub.13, R.sub.17, R.sub.28, R.sub.30, R.sub.34, R.sub.39, R.sub.58, R.sub.62, R.sub.67, R.sub.68=are independently C, U or absent; [0541] R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.10, R.sub.19, R.sub.20, R.sub.23, R.sub.52=are independently G or absent; [0542] R.sub.33, R.sub.36=are independently G, U or absent; [0543] R.sub.8, R.sub.21, R.sub.54, R.sub.69=are independently U or absent; [0544] [R.sub.47].sub.x=N or absent;

[0545] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Arginine TREM Consensus Sequence

[0546] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I A.sub.RG (SEQ ID NO: 565),

[0547] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Arg is: [0548] R.sub.57=A or absent; [0549] R.sub.9,R.sub.27=are independently A, C, G or absent; [0550] R.sub.1,R.sub.2,R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.11,R.sub.12,R.sub.16,R.sub.21,R.sub.22,R.sub.23,R.sub.25,R.sub.26,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.33,R.sub.34,R.sub.37,R.sub.42,R.sub.44,R.sub.45, R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.51,R.sub.58,R.sub.62,R.sub.63,R.sub.64,R.sub.68,R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.70,R.sub.71=are independently N or absent; [0551] R.sub.13,R.sub.17,R.sub.41=are independently A, C, U or absent; [0552] R.sub.19,R.sub.20,R.sub.24,R.sub.40,R.sub.56=are independently A, G or absent; [0553] R.sub.14,R.sub.15,R.sub.72=are independently A, G, U or absent; [0554] R.sub.18=A, U or absent; [0555] R.sub.38=C or absent; [0556] R.sub.35,R.sub.43,R.sub.61=are independently C, G, U or absent; [0557] R.sub.28,R.sub.55,R.sub.59,R.sub.60=are independently C, U or absent; [0558] R.sub.0,R.sub.10,R.sub.52=are independently G or absent; [0559] R.sub.8,R.sub.39=are independently G, U or absent; [0560] R.sub.36,R.sub.53,R.sub.54=are independently U or absent; [0561] [R.sub.47].sub.x=N or absent;

[0562] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0563] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II A.sub.RG (SEQ ID NO: 566),

[0564] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

[0565] wherein R is a ribonucleotide residue and the consensus for Arg is: [0566] R.sub.18=absent; [0567] R.sub.24,R.sub.57=are independently A or absent; [0568] R.sub.41=A, C or absent; [0569] R.sub.3,R.sub.7,R.sub.34,R.sub.50=are independently A, C, G or absent; [0570] R.sub.2,R.sub.5,R.sub.6,R.sub.12,R.sub.26,R.sub.32,R.sub.37,R.sub.44,R.sub.58,R.sub.66,R.sub.67,R.sub.68,R.sub.70=are independently N or absent; [0571] R.sub.49,R.sub.71=are independently A, C, U or absent; [0572] R.sub.1,R.sub.15,R.sub.19,R.sub.25,R.sub.27,R.sub.40,R.sub.45,R.sub.46,R.sub.56,R.sub.72=are independently A, G or absent; [0573] R.sub.14,R.sub.29,R.sub.63=are independently A, G, U or absent; [0574] R.sub.16,R.sub.21=are independently A, U or absent; [0575] R.sub.38,R.sub.61=are independently C or absent; [0576] R.sub.33,R.sub.48=are independently C, G or absent; [0577] R.sub.4,R.sub.9,R.sub.11,R.sub.43,R.sub.62,R.sub.64,R.sub.69=are independently C, G, U or absent; [0578] R.sub.13,R.sub.22,R.sub.28,R.sub.30,R.sub.31,R.sub.35,R.sub.55,R.sub.60,R.sub.65=are independently C, U or absent; [0579] R.sub.0,R.sub.10,R.sub.20,R.sub.23,R.sub.51,R.sub.52=are independently G or absent; [0580] R.sub.8,R.sub.39,R.sub.42=are independently G, U or absent; [0581] R.sub.17,R.sub.36,R.sub.53,R.sub.54,R.sub.59=are independently U or absent; [0582] [R.sub.47].sub.x=N or absent;

[0583] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0584] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III A.sub.RG (SEQ ID NO: 567),

[0585] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

[0586] wherein R is a ribonucleotide residue and the consensus for Arg is: [0587] R.sub.18=is absent; [0588] R.sub.15,R.sub.21,R.sub.24,R.sub.41,R.sub.57=are independently A or absent; [0589] R.sub.34,R.sub.44=are independently A, C or absent; [0590] R.sub.3,R.sub.5,R.sub.58=are independently A, C, G or absent; [0591] R.sub.2,R.sub.6,R.sub.66,R.sub.70=are independently N or absent; [0592] R.sub.37,R.sub.49=are independently A, C, U or absent; [0593] R.sub.1,R.sub.25,R.sub.29,R.sub.40,R.sub.45,R.sub.46,R.sub.50=are independently A, G or absent; [0594] R.sub.14,R.sub.63,R.sub.68=are independently A, G, U or absent; [0595] R.sub.16=A, U or absent; [0596] R.sub.38,R.sub.61=are independently C or absent; [0597] R.sub.7,R.sub.11,R.sub.12,R.sub.26,R.sub.48=are independently C, G or absent; [0598] R.sub.64,R.sub.67,R.sub.69=are independently C, G, U or absent; [0599] R.sub.4,R.sub.13,R.sub.22,R.sub.28,R.sub.30,R.sub.31,R.sub.35,R.sub.43,R.sub.58,R.sub.60,R.sub.62,R.sub.68,R.sub.71=are independently C, U or absent; [0600] R.sub.0,R.sub.10,R.sub.19,R.sub.20,R.sub.23,R.sub.27,R.sub.33,R.sub.51,R.sub.52,R.sub.56,R.sub.72=are independently G or absent; [0601] R.sub.8,R.sub.9,R.sub.32,R.sub.39,R.sub.42=are independently G, U or absent; [0602] R.sub.17,R.sub.36,R.sub.53,R.sub.54,R.sub.59=are independently U or absent; [0603] [R.sub.47].sub.x=N or absent;

[0604] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Asparagine TREM Consensus Sequence

[0605] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.ASN (SEQ ID NO: 568),

[0606] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Asn is: [0607] R.sub.0,R.sub.18=are absent; [0608] R.sub.41=A or absent; [0609] R.sub.14,R.sub.48,R.sub.56=are independently A, C, G or absent; [0610] R.sub.2,R.sub.4,R.sub.5,R.sub.6,R.sub.12,R.sub.17,R.sub.26,R.sub.29,R.sub.30,R.sub.31,R.sub.44,R.sub.45,R.sub.46,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.65,R.sub.66,R.sub.67,R.sub.68,R.sub.70,R.sub.71=are independently N or absent; [0611] R.sub.11,R.sub.13,R.sub.22,R.sub.42,R.sub.55,R.sub.59=are independently A, C, U or absent; [0612] R.sub.9,R.sub.15,R.sub.24,R.sub.27,R.sub.34,R.sub.37,R.sub.51,R.sub.72=are independently A, G or absent; [0613] R.sub.1,R.sub.7,R.sub.25,R.sub.69=are independently A, G, U or absent; [0614] R.sub.40,R.sub.57=are independently A, U or absent; [0615] R.sub.60=C or absent; [0616] R.sub.33=C, G or absent; [0617] R.sub.21,R.sub.32,R.sub.43,R.sub.64=are independently C, G, U or absent; [0618] R.sub.3,R.sub.16,R.sub.28,R.sub.35,R.sub.36,R.sub.61=are independently C, U or absent; [0619] R.sub.10,R.sub.19,R.sub.20,R.sub.52=are independently G or absent; [0620] R.sub.54=G, U or absent; [0621] R.sub.8,R.sub.23,R.sub.38,R.sub.39,R.sub.53=are independently U or absent; [0622] [R.sub.47].sub.x=N or absent;

[0623] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0624] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.ASN (SEQ ID NO: 569),

[0625] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Asn is: [0626] R.sub.0,R.sub.18=are absent [0627] R.sub.24,R.sub.41,R.sub.46,R.sub.62=are independently A or absent; [0628] R.sub.59=A, C or absent; [0629] R.sub.14,R.sub.56,R.sub.66=are independently A, C, G or absent; [0630] R.sub.17,R.sub.29=are independently N or absent; [0631] R.sub.11,R.sub.26,R.sub.42,R.sub.55=are independently A, C, U or absent; [0632] R.sub.1,R.sub.9,R.sub.12,R.sub.15,R.sub.25,R.sub.34,R.sub.37,R.sub.48,R.sub.51,R.sub.67,R.sub.68,R.sub.69,R.sub.70,R.sub.72=are independently A, G or absent; [0633] R.sub.44,R.sub.45,R.sub.58=are independently A, G, U or absent; [0634] R.sub.40,R.sub.57=are independently A, U or absent; [0635] R.sub.5,R.sub.28,R.sub.60=are independently C or absent; [0636] R.sub.33,R.sub.65=are independently C, G or absent; [0637] R.sub.21,R.sub.43,R.sub.71=are independently C, G, U or absent; [0638] R.sub.3,R.sub.6,R.sub.13,R.sub.22,R.sub.32,R.sub.35,R.sub.36,R.sub.61,R.sub.63,R.sub.64=are independently C, U or absent; [0639] R.sub.7,R.sub.10,R.sub.19,R.sub.20,R.sub.27,R.sub.49,R.sub.52=are independently G or absent; [0640] R.sub.54=G, U or absent; [0641] R.sub.2,R.sub.4,R.sub.5,R.sub.16,R.sub.23,R.sub.30,R.sub.31,R.sub.38,R.sub.39,R.sub.50,R.sub.53=are independently U or absent; [0642] [R.sub.47].sub.x=N or absent;

[0643] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0644] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.ASN (SEQ ID NO: 570),

[0645] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Asn is: [0646] R.sub.0,R.sub.18=are absent [0647] R.sub.24,R.sub.40,R.sub.41,R.sub.46,R.sub.62=are independently A or absent; [0648] R.sub.59=A, C or absent; [0649] R.sub.14,R.sub.56,R.sub.66=are independently A, C, G or absent; [0650] R.sub.11,R.sub.26,R.sub.42,R.sub.55=are independently A, C, U or absent; [0651] R.sub.1,R.sub.9,R.sub.12,R.sub.15,R.sub.34,R.sub.37,R.sub.48,R.sub.51,R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently A, G or absent; [0652] R.sub.44,R.sub.45,R.sub.58=are independently A, G, U or absent; [0653] R.sub.57=A, U or absent; [0654] R.sub.5,R.sub.28,R.sub.60=are independently C or absent; [0655] R.sub.33,R.sub.65=are independently C, G or absent; [0656] R.sub.17,R.sub.21,R.sub.29=are independently C, G, U or absent; [0657] R.sub.3,R.sub.6,R.sub.13,R.sub.22,R.sub.32,R.sub.35,R.sub.36,R.sub.43,R.sub.61,R.sub.63,R.sub.64,R.sub.71=are independently C, U or absent; [0658] R.sub.7,R.sub.10,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.49,R.sub.52,R.sub.72=are independently G or absent; [0659] R.sub.54=G, U or absent; [0660] R.sub.2,R.sub.4,R.sub.8,R.sub.16,R.sub.23,R.sub.30,R.sub.31,R.sub.38,R.sub.39,R.sub.50,R.sub.53=are independently U or absent; [0661] [R.sub.47].sub.x=N or absent;

[0662] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Aspartate TREM Consensus Sequence

[0663] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I ASP (SEQ ID NO: 571),

[0664] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Asp is: [0665] R.sub.0=absent [0666] R.sub.24,R.sub.71=are independently A, C or absent; [0667] R.sub.33,R.sub.46=are independently A, C, G or absent; [0668] R.sub.2,R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.12,R.sub.16,R.sub.22,R.sub.26,R.sub.29,R.sub.31,R.sub.32,R.sub.44,R.sub.48,R.sub.49,R.sub.58,R.sub.63,R.sub.64,R.sub.66,R.sub.67,R.sub.68,R.sub.69=are independently N or absent; [0669] R.sub.13,R.sub.21,R.sub.34,R.sub.41,R.sub.57,R.sub.65=are independently A, C, U or absent; [0670] R.sub.9,R.sub.10,R.sub.14,R.sub.15,R.sub.20,R.sub.27,R.sub.37,R.sub.40,R.sub.51,R.sub.56,R.sub.72=are independently A, G or absent; [0671] R.sub.7,R.sub.25,R.sub.42=are independently A, G, U or absent; [0672] R.sub.39=C or absent; [0673] R.sub.50,R.sub.62=are independently C, G or absent; [0674] R.sub.30,R.sub.43,R.sub.45,R.sub.55,R.sub.70=are independently C, G, U or absent; [0675] R.sub.8,R.sub.11,R.sub.17,R.sub.18,R.sub.28,R.sub.35,R.sub.53,R.sub.59,R.sub.60,R.sub.61=are independently C, U or absent; [0676] R.sub.19,R.sub.52=are independently G or absent; [0677] R.sub.1=G, U or absent; [0678] R.sub.23,R.sub.36,R.sub.38,R.sub.54=are independently U or absent; [0679] [R.sub.47].sub.x=N or absent;

[0680] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0681] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II ASP (SEQ ID NO: 572),

[0682] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Asp is: [0683] R.sub.0,R.sub.17,R.sub.18,R.sub.23=are independently absent; [0684] R.sub.9,R.sub.40=are independently A or absent; [0685] R.sub.24,R.sub.71=are independently A, C or absent; [0686] R.sub.67,R.sub.68=are independently A, C, G or absent; [0687] R.sub.2,R.sub.6,R.sub.66=are independently N or absent; [0688] R.sub.57,R.sub.63=are independently A, C, U or absent; [0689] R.sub.10,R.sub.14,R.sub.27,R.sub.33,R.sub.37,R.sub.44,R.sub.46,R.sub.51,R.sub.56,R.sub.64,R.sub.72=are independently A, G or absent; [0690] R.sub.7,R.sub.12,R.sub.26,R.sub.65=are independently A, U or absent; [0691] R.sub.39,R.sub.61,R.sub.62=are independently C or absent; [0692] R.sub.3,R.sub.31,R.sub.45,R.sub.70=are independently C, G or absent; [0693] R.sub.4,R.sub.5,R.sub.29,R.sub.43,R.sub.55=are independently C, G, U or absent; [0694] R.sub.8,R.sub.11,R.sub.13,R.sub.30,R.sub.32,R.sub.34,R.sub.35,R.sub.41,R.sub.48,R.sub.53,R.sub.59,R.sub.60=are independently C, U or absent; [0695] R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.42,R.sub.50,R.sub.52=are independently G or absent; [0696] R.sub.1,R.sub.22,R.sub.49,R.sub.58,R.sub.69=are independently G, U or absent; [0697] R.sub.16,R.sub.21,R.sub.28,R.sub.36,R.sub.38,R.sub.54=are independently U or absent; [0698] [R.sub.47].sub.x=N or absent;

[0699] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0700] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III ASP (SEQ ID NO: 573),

[0701] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Asp is: [0702] R.sub.0,R.sub.17,R.sub.18,R.sub.23=are absent [0703] R.sub.9,R.sub.12,R.sub.40,R.sub.65,R.sub.71=are independently A or absent; [0704] R.sub.2,R.sub.24,R.sub.57=are independently A, C or absent; [0705] R.sub.6,R.sub.14,R.sub.27,R.sub.46,R.sub.51,R.sub.56,R.sub.64,R.sub.67,R.sub.68=are independently A, G or absent; [0706] R.sub.3,R.sub.31,R.sub.35,R.sub.39,R.sub.61,R.sub.62=are independently C or absent; [0707] R.sub.66=C, G or absent; [0708] R.sub.5,R.sub.8,R.sub.29,R.sub.30,R.sub.32,R.sub.34,R.sub.41,R.sub.43,R.sub.48,R.sub.55,R.sub.59,R.sub.60,R.sub.63=are independently C, U or absent; [0709] R.sub.10,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.33,R.sub.37,R.sub.42,R.sub.44,R.sub.45,R.sub.49,R.sub.50,R.sub.52,R.sub.69,R.sub.70,R.sub.72=are independently G or absent; [0710] R.sub.22,R.sub.58=are independently G, U or absent; [0711] R.sub.1,R.sub.4,R.sub.7,R.sub.11,R.sub.13,R.sub.16,R.sub.21,R.sub.26,R.sub.28,R.sub.36,R.sub.38,R.sub.53,R.sub.54=are independently U or absent; [0712] [R.sub.47].sub.x=N or absent;

[0713] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Cysteine TREM Consensus Sequence

[0714] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.CYS (SEQ ID NO: 574),

[0715] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Cys is: [0716] R.sub.0=absent [0717] R.sub.14,R.sub.39,R.sub.57=are independently A or absent; [0718] R.sub.41=A, C or absent; [0719] R.sub.10,R.sub.15,R.sub.27,R.sub.33,R.sub.62=are independently A, C, G or absent; [0720] R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.12,R.sub.13,R.sub.16,R.sub.24,R.sub.26,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.34,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.58,R.sub.63,R.sub.64,R.sub.66, [0721] R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [0722] R.sub.65=A, C, U or absent; [0723] R.sub.9,R.sub.25,R.sub.37,R.sub.40,R.sub.52,R.sub.56=are independently A, G or absent; [0724] R.sub.7,R.sub.20,R.sub.51=are independently A, G, U or absent; [0725] R.sub.18,R.sub.38,R.sub.55=are independently C or absent; [0726] R.sub.2=C, G or absent; [0727] R.sub.21,R.sub.28,R.sub.43,R.sub.50=are independently C, G, U or absent; [0728] R.sub.11,R.sub.22,R.sub.23,R.sub.35,R.sub.36,R.sub.59,R.sub.60,R.sub.61,R.sub.71,R.sub.72=are independently C, U or absent; [0729] R.sub.1,R.sub.19=are independently G or absent; [0730] R.sub.17=G, U or absent; [0731] R.sub.8,R.sub.53,R.sub.54=are independently U or absent; [0732] [R.sub.47].sub.x=N or absent;

[0733] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0734] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.CYS (SEQ ID NO: 575), [0735] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72
wherein R is a ribonucleotide residue and the consensus for Cys is: [0736] R.sub.0,R.sub.18,R.sub.23=are absent; [0737] R.sub.14,R.sub.24,R.sub.26,R.sub.29,R.sub.39,R.sub.41,R.sub.45,R.sub.57=are independently A or absent; [0738] R.sub.44=A, C or absent; [0739] R.sub.27,R.sub.62=are independently A, C, G or absent; [0740] R.sub.16=A, C, G, U or absent; [0741] R.sub.30,R.sub.70=are independently A, C, U or absent; [0742] R.sub.5,R.sub.7,R.sub.9,R.sub.25,R.sub.34,R.sub.37,R.sub.40,R.sub.46,R.sub.52,R.sub.56,R.sub.58,R.sub.66=are independently A, G or absent; [0743] R.sub.20,R.sub.51=are independently A, G, U or absent; [0744] R.sub.35,R.sub.38,R.sub.43,R.sub.55,R.sub.69=are independently C or absent; [0745] R.sub.2,R.sub.4,R.sub.15=are independently C, G or absent; [0746] R.sub.13=C, G, U or absent; [0747] R.sub.6,R.sub.11,R.sub.28,R.sub.36,R.sub.48,R.sub.49,R.sub.50,R.sub.60,R.sub.61,R.sub.67,R.sub.68,R.sub.71,R.sub.72=are independently C, U or absent; [0748] R.sub.1,R.sub.3,R.sub.10,R.sub.19,R.sub.33,R.sub.63=are independently G or absent; [0749] R.sub.8,R.sub.17,R.sub.21,R.sub.64=are independently G, U or absent; [0750] R.sub.12,R.sub.22,R.sub.31,R.sub.32,R.sub.42,R.sub.53,R.sub.54,R.sub.65=are independently U or absent; [0751] R.sub.59=U, or absent; [0752] [R.sub.47].sub.x=N or absent;

[0753] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0754] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.CYS (SEQ ID NO: 576),

[0755] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Cys is: [0756] R.sub.0,R.sub.18,R.sub.23=are absent [0757] R.sub.14,R.sub.24,R.sub.26,R.sub.29,R.sub.34,R.sub.39,R.sub.41,R.sub.45,R.sub.57,R.sub.58=are independently A or absent; [0758] R.sub.44,R.sub.70=are independently A, C or absent; [0759] R.sub.62=A, C, G or absent; [0760] R.sub.16=N or absent; [0761] R.sub.5,R.sub.7,R.sub.9,R.sub.20,R.sub.40,R.sub.46,R.sub.51,R.sub.52,R.sub.56,R.sub.66=are independently A, G or absent; [0762] R.sub.28,R.sub.35,R.sub.38,R.sub.43,R.sub.55,R.sub.67,R.sub.69=are independently C or absent; [0763] R.sub.4,R.sub.15=are independently C, G or absent; [0764] R.sub.6,R.sub.11,R.sub.3,R.sub.30,R.sub.48,R.sub.49,R.sub.50,R.sub.60,R.sub.61,R.sub.68,R.sub.71,R.sub.72=are independently C, U or absent; [0765] R.sub.1,R.sub.2,R.sub.3,R.sub.10,R.sub.19,R.sub.25,R.sub.27,R.sub.33,R.sub.37,R.sub.63=are independently G or absent; [0766] R.sub.8,R.sub.21,R.sub.64=are independently G, U or absent; [0767] R.sub.12,R.sub.17,R.sub.22,R.sub.31,R.sub.32,R.sub.36,R.sub.42,R.sub.53,R.sub.54, R.sub.59,R.sub.65=are independently U or absent; [0768] [R.sub.47].sub.x=N or absent;

[0769] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Glutamine TREM Consensus Sequence

[0770] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.GLN(SEQ ID NO: 577),

[0771] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Gln is: [0772] R.sub.0,R.sub.18=are absent; [0773] R.sub.14,R.sub.24,R.sub.57=are independently A or absent; [0774] R.sub.9,R.sub.26,R.sub.27,R.sub.33,R.sub.56=are independently A, C, G or absent; [0775] R.sub.2,R.sub.4,R.sub.5,R.sub.6,R.sub.12,R.sub.13,R.sub.16,R.sub.21,R.sub.22,R.sub.2,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.34,R.sub.41,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [0776] R.sub.17,R.sub.23,R.sub.43,R.sub.65,R.sub.71=are independently A, C, U or absent; [0777] R.sub.15,R.sub.40,R.sub.51,R.sub.52=are independently A, G or absent; [0778] R.sub.1,R.sub.7,R.sub.72=are independently A, G, U or absent; [0779] R.sub.3,R.sub.11,R.sub.37,R.sub.60,R.sub.64=are independently C, G, U or absent; [0780] R.sub.28,R.sub.35,R.sub.55,R.sub.59,R.sub.61=are independently C, U or absent; [0781] R.sub.10,R.sub.19,R.sub.20=are independently G or absent; [0782] R.sub.39=G, U or absent; [0783] R.sub.8,R.sub.36,R.sub.38,R.sub.53,R.sub.54=are independently U or absent; [0784] [R.sub.47].sub.x=N or absent;

[0785] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0786] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.GLN (SEQ ID NO: 578),

[0787] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Gln is: [0788] R.sub.0,R.sub.18,R.sub.23=are absent [0789] R.sub.14,R.sub.24,R.sub.57=are independently A or absent; [0790] R.sub.17,R.sub.71=are independently A, C or absent; [0791] R.sub.25,R.sub.26,R.sub.33,R.sub.44,R.sub.46,R.sub.56,R.sub.69=are independently A, C, G or absent; [0792] R.sub.4,R.sub.5,R.sub.12,R.sub.22,R.sub.29,R.sub.30,R.sub.48,R.sub.49,R.sub.63,R.sub.67,R.sub.68=are independently N or absent; [0793] R.sub.31,R.sub.43,R.sub.62,R.sub.65,R.sub.70=are independently A, C, U or absent; [0794] R.sub.15,R.sub.27,R.sub.34,R.sub.40,R.sub.41,R.sub.51,R.sub.52=are independently A, G or absent; [0795] R.sub.2,R.sub.7,R.sub.21,R.sub.45,R.sub.50,R.sub.58,R.sub.66,R.sub.72=are independently A, G, U or absent; [0796] R.sub.3,R.sub.13,R.sub.32,R.sub.37,R.sub.42,R.sub.60,R.sub.64=are independently C, G, U or absent; [0797] R.sub.6,R.sub.11,R.sub.28,R.sub.35,R.sub.55,R.sub.59,R.sub.61=are independently C, U or absent; [0798] R.sub.9,R.sub.10,R.sub.19,R.sub.20=are independently G or absent; [0799] R.sub.1,R.sub.16,R.sub.39=are independently G, U or absent; [0800] R.sub.8,R.sub.36,R.sub.38,R.sub.53,R.sub.54=are independently U or absent; [0801] [R.sub.47].sub.x=N or absent;

[0802] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0803] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.GLN (SEQ ID NO: 579),

[0804] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Gln is: [0805] R.sub.0,R.sub.18,R.sub.23=are absent [0806] R.sub.14,R.sub.24,R.sub.41,R.sub.57=are independently A or absent; [0807] R.sub.17,R.sub.71=are independently A, C or absent; [0808] R.sub.5,R.sub.25,R.sub.26,R.sub.46,R.sub.56,R.sub.69=are independently A, C, G or absent; [0809] R.sub.4,R.sub.22,R.sub.29,R.sub.30,R.sub.48,R.sub.49,R.sub.63,R.sub.68=are independently N or absent; [0810] R.sub.43,R.sub.62,R.sub.65,R.sub.70=are independently A, C, U or absent; [0811] R.sub.15,R.sub.27,R.sub.33,R.sub.34,R.sub.40,R.sub.51,R.sub.52=are independently A, G or absent; [0812] R.sub.2,R.sub.7,R.sub.12,R.sub.45,R.sub.50,R.sub.58,R.sub.66=are independently A, G, U or absent; [0813] R.sub.31=A, U or absent; [0814] R.sub.32,R.sub.44,R.sub.60=are independently C, G or absent; [0815] R.sub.3,R.sub.13,R.sub.37,R.sub.42,R.sub.64,R.sub.67=are independently C, G, U or absent; [0816] R.sub.6,R.sub.11,R.sub.28,R.sub.35,R.sub.55,R.sub.59,R.sub.61=are independently C, U or absent; [0817] R.sub.9,R.sub.10,R.sub.19,R.sub.20=are independently G or absent; [0818] R.sub.1,R.sub.21,R.sub.39,R.sub.72=are independently G, U or absent; [0819] R.sub.8,R.sub.16,R.sub.36,R.sub.38,R.sub.53,R.sub.54=are independently U or absent; [0820] [R.sub.47].sub.x=N or absent;

[0821] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Glutamate TREM Consensus Sequence

[0822] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.GLU (SEQ ID NO: 580),

[0823] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Glu is: [0824] R.sub.0=absent; [0825] R.sub.34,R.sub.43,R.sub.68,R.sub.69=are independently A, C, G or absent; [0826] R.sub.1,R.sub.2,R.sub.5,R.sub.6,R.sub.9,R.sub.12,R.sub.16,R.sub.20,R.sub.21,R.sub.26,R.sub.27,R.sub.29,R.sub.3,R.sub.31,R.sub.32,R.sub.33,R.sub.41,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.50,R.sub.51,R.sub.55,R.sub.63,R.sub.64,R.sub.65,R.sub.66,R.sub.70,R.sub.71=are independently N or absent; [0827] R.sub.13,R.sub.17,R.sub.23,R.sub.61=are independently A, C, U or absent; [0828] R.sub.10,R.sub.14,R.sub.24,R.sub.40,R.sub.52,R.sub.56=are independently A, G or absent; [0829] R.sub.7,R.sub.15,R.sub.25,R.sub.67,R.sub.72=are independently A, G, U or absent; [0830] R.sub.11,R.sub.57=are independently A, U or absent; [0831] R.sub.39=C, G or absent; [0832] R.sub.3,R.sub.4,R.sub.22,R.sub.42,R.sub.49,R.sub.55,R.sub.62=are independently C, G, U or absent; [0833] R.sub.18,R.sub.28,R.sub.35,R.sub.37,R.sub.53,R.sub.59,R.sub.60=are independently C, U or absent; R.sub.19=G or absent; [0834] R.sub.8,R.sub.36,R.sub.38,R.sub.54=are independently U or absent; [0835] [R.sub.47].sub.x=N or absent;

[0836] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0837] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.GLU (SEQ ID NO: 581),

[0838] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Glu is: [0839] R.sub.0,R.sub.18,R.sub.23=are absent [0840] R.sub.17,R.sub.40=are independently A or absent; [0841] R.sub.26,R.sub.27,R.sub.34,R.sub.43,R.sub.68,R.sub.69,R.sub.71=are independently A, C, G or absent; [0842] R.sub.1,R.sub.2,R.sub.5,R.sub.12,R.sub.21,R.sub.31,R.sub.33,R.sub.41,R.sub.45,R.sub.48,R.sub.51,R.sub.58,R.sub.66,R.sub.70=are independently N or absent; [0843] R.sub.44,R.sub.61=are independently A, C, U or absent; [0844] R.sub.9,R.sub.14,R.sub.24,R.sub.25,R.sub.52,R.sub.56,R.sub.63=are independently A, G or absent; [0845] R.sub.7,R.sub.15,R.sub.46,R.sub.50,R.sub.67,R.sub.72=are independently A, G, U or absent; [0846] R.sub.29,R.sub.57=are independently A, U or absent; [0847] R.sub.60=C or absent; [0848] R.sub.39=C, G or absent; [0849] R.sub.3,R.sub.6,R.sub.20,R.sub.30,R.sub.32,R.sub.42,R.sub.55,R.sub.62,R.sub.65=are independently C, G, U or absent; [0850] R.sub.4,R.sub.8,R.sub.16,R.sub.28,R.sub.35,R.sub.37,R.sub.49,R.sub.53,R.sub.59=are independently C, U or absent; [0851] R.sub.10,R.sub.19=are independently G or absent; [0852] R.sub.22,R.sub.64=are independently G, U or absent; [0853] R.sub.11,R.sub.13,R.sub.36,R.sub.38,R.sub.54=are independently U or absent; [0854] [R.sub.47].sub.x=N or absent;

[0855] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0856] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.GLU (SEQ ID NO: 582),

[0857] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Glu is: [0858] R.sub.0,R.sub.17,R.sub.18,R.sub.23=are absent [0859] R.sub.14,R.sub.27,R.sub.40,R.sub.71=are independently A or absent; [0860] R.sub.44=A, C or absent; [0861] R.sub.43=A, C, G or absent; [0862] R.sub.1,R.sub.31,R.sub.33,R.sub.45,R.sub.51,R.sub.66=are independently N or absent; [0863] R.sub.21,R.sub.41=are independently A, C, U or absent; [0864] R.sub.7,R.sub.24,R.sub.25,R.sub.50,R.sub.52,R.sub.56,R.sub.63,R.sub.68,R.sub.70=are independently A, G or absent; [0865] R.sub.5,R.sub.46=are independently A, G, U or absent; [0866] R.sub.29,R.sub.57,R.sub.67,R.sub.72=are independently A, U or absent; [0867] R.sub.2,R.sub.39,R.sub.60=are independently C or absent; [0868] R.sub.3,R.sub.12,R.sub.20,R.sub.26,R.sub.34,R.sub.69=are independently C, G or absent; [0869] R.sub.6,R.sub.30,R.sub.42,R.sub.48,R.sub.69=are independently C, G, U or absent; [0870] R.sub.4,R.sub.16,R.sub.28,R.sub.35,R.sub.37,R.sub.49,R.sub.53,R.sub.55,R.sub.58,R.sub.61,R.sub.62=are independently C, U or absent; [0871] R.sub.9,R.sub.10,R.sub.19,R.sub.64=are independently G or absent; [0872] R.sub.15,R.sub.22,R.sub.32=are independently G, U or absent; [0873] R.sub.8,R.sub.11,R.sub.13,R.sub.36,R.sub.38,R.sub.54,R.sub.59=are independently U or absent; [0874] [R.sub.47].sub.x=N or absent;

[0875] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Glycine TREM Consensus Sequence

[0876] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.GLY(SEQ ID NO: 583),

[0877] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Gly is: [0878] R.sub.0=absent; [0879] R.sub.24=A or absent; [0880] R.sub.3,R.sub.9,R.sub.40,R.sub.50,R.sub.51=are independently A, C, G or absent; [0881] R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.12,R.sub.16,R.sub.21,R.sub.22,R.sub.26,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.33,R.sub.34,R.sub.41,R.sub.42,R.sub.43,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.58,R.sub.63,R.sub.64,R.sub.65,R.sub.66,R.sub.67,R.sub.68=are independently N or absent; [0882] R.sub.59=A, C, U or absent; [0883] R.sub.1,R.sub.10,R.sub.14,R.sub.15,R.sub.27,R.sub.56=are independently A, G or absent; [0884] R.sub.20,R.sub.25=are independently A, G, U or absent; [0885] R.sub.57,R.sub.72=are independently A, U or absent; [0886] R.sub.38,R.sub.39,R.sub.60=are independently C or absent; [0887] R.sub.52=C, G or absent; [0888] R.sub.2,R.sub.19,R.sub.37,R.sub.54,R.sub.55,R.sub.61,R.sub.62,R.sub.69,R.sub.70=are independently C, G, U or absent; [0889] R.sub.11,R.sub.13,R.sub.17,R.sub.28,R.sub.35,R.sub.36,R.sub.71=are independently C, U or absent; [0890] R.sub.8,R.sub.18,R.sub.23,R.sub.53=are independently U or absent; [0891] [R.sub.47].sub.x=N or absent;

[0892] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0893] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.GLY (SEQ ID NO: 584),

[0894] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65- R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Gly is: [0895] R.sub.0,R.sub.18,R.sub.23=are absent [0896] R.sub.24,R.sub.27,R.sub.40,R.sub.72=are independently A or absent; [0897] R.sub.26=A, C or absent; [0898] R.sub.3,R.sub.7,R.sub.68=are independently A, C, G or absent; [0899] R.sub.5,R.sub.30,R.sub.41,R.sub.42,R.sub.44,R.sub.49,R.sub.67=are independently A, C, G, U or absent; [0900] R.sub.31,R.sub.32,R.sub.34=are independently A, C, U or absent; [0901] R.sub.9,R.sub.10,R.sub.14,R.sub.15,R.sub.33,R.sub.50,R.sub.56=are independently A, G or absent; [0902] R.sub.12,R.sub.16,R.sub.22,R.sub.25,R.sub.29,R.sub.46=are independently A, G, U or absent; [0903] R.sub.57=A, U or absent; [0904] R.sub.17,R.sub.38,R.sub.39,R.sub.60,R.sub.61,R.sub.71=are independently C or absent; [0905] R.sub.6,R.sub.52,R.sub.64,R.sub.66=are independently C, G or absent; [0906] R.sub.2,R.sub.4,R.sub.37,R.sub.48,R.sub.55,R.sub.65=are independently C, G, U or absent; [0907] R.sub.13,R.sub.35,R.sub.43,R.sub.62,R.sub.69=are independently C, U or absent; [0908] R.sub.1,R.sub.19,R.sub.20,R.sub.51,R.sub.70=are independently G or absent; [0909] R.sub.21,R.sub.45,R.sub.63=are independently G, U or absent; [0910] R.sub.8,R.sub.11,R.sub.28,R.sub.36,R.sub.53,R.sub.54,R.sub.58,R.sub.59=are independently U or absent; [0911] [R.sub.47].sub.x=N or absent;

[0912] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0913] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.GLY (SEQ ID NO: 585),

[0914] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Gly is: [0915] R.sub.0,R.sub.18,R.sub.23=are absent [0916] R.sub.24,R.sub.27,R.sub.40,R.sub.72=are independently A or absent; [0917] R.sub.26=A, C or absent; [0918] R.sub.3,R.sub.7,R.sub.49,R.sub.68=are independently A, C, G or absent; [0919] R.sub.5,R.sub.30,R.sub.41,R.sub.44,R.sub.67=are independently N or absent; [0920] R.sub.31,R.sub.32,R.sub.34=are independently A, C, U or absent; [0921] R.sub.9,R.sub.10,R.sub.14,R.sub.15,R.sub.33,R.sub.50,R.sub.56=are independently A, G or absent; [0922] R.sub.12,R.sub.25,R.sub.29,R.sub.42,R.sub.46=are independently A, G, U or absent; [0923] R.sub.16,R.sub.57=are independently A, U or absent; [0924] R.sub.17,R.sub.38,R.sub.39,R.sub.60,R.sub.61,R.sub.71=are independently C or absent; [0925] R.sub.6,R.sub.52,R.sub.64,R.sub.66=are independently C, G or absent; [0926] R.sub.37,R.sub.48,R.sub.65=are independently C, G, U or absent; [0927] R.sub.2,R.sub.4,R.sub.13,R.sub.35,R.sub.43,R.sub.55,R.sub.62,R.sub.69=are independently C, U or absent; [0928] R.sub.1,R.sub.19,R.sub.20,R.sub.51,R.sub.70=are independently G or absent; [0929] R.sub.21,R.sub.22,R.sub.45,R.sub.63=are independently G, U or absent; [0930] R.sub.8,R.sub.11,R.sub.28,R.sub.36,R.sub.53,R.sub.54,R.sub.58,R.sub.59=are independently U or absent; [0931] [R.sub.47].sub.x=N or absent;

[0932] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Histidine TREM Consensus Sequence

[0933] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.HIS (SEQ ID NO: 586),

[0934] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.1-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for His is: [0935] R.sub.23=absent; [0936] R.sub.14,R.sub.24,R.sub.57=are independently A or absent; [0937] R.sub.72=A, C or absent; [0938] R.sub.9,R.sub.27,R.sub.43,R.sub.48,R.sub.69=are independently A, C, G or absent; [0939] R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.12,R.sub.25,R.sub.26,R.sub.29,R.sub.30,R.sub.31,R.sub.34,R.sub.42,R.sub.45,R.sub.46,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.66,R.sub.67,R.sub.68=are independently N or absent; [0940] R.sub.13,R.sub.21,R.sub.41,R.sub.44,R.sub.65=are independently A, C, U or absent; [0941] R.sub.40,R.sub.51,R.sub.56,R.sub.70=are independently A, G or absent; [0942] R.sub.7,R.sub.32=are independently A, G, U or absent; [0943] R.sub.55,R.sub.60=are independently C or absent; [0944] R.sub.11,R.sub.16,R.sub.33,R.sub.64=are independently C, G, U or absent; [0945] R.sub.2,R.sub.17,R.sub.22,R.sub.28,R.sub.35,R.sub.53,R.sub.59,R.sub.61,R.sub.71=are independently C, U or absent; [0946] R.sub.1,R.sub.10,R.sub.15,R.sub.19,R.sub.2,R.sub.37,R.sub.39,R.sub.52=are independently G or absent; R.sub.0=G, U or absent; [0947] R.sub.8,R.sub.18,R.sub.36,R.sub.38,R.sub.54=are independently U or absent; [0948] [R.sub.47].sub.x=N or absent;

[0949] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0950] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.HIS (SEQ ID NO: 587),

[0951] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.1-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for His is: [0952] R.sub.0,R.sub.17,R.sub.18,R.sub.23=are absent; [0953] R.sub.7,R.sub.12,R.sub.14,R.sub.24,R.sub.27,R.sub.45,R.sub.57,R.sub.58,R.sub.63,R.sub.67,R.sub.72=are independently A or absent; [0954] R.sub.3=A, C, U or absent; [0955] R.sub.4,R.sub.43,R.sub.56,R.sub.70=are independently A, G or absent; [0956] R.sub.49=A, U or absent; [0957] R.sub.2,R.sub.28,R.sub.30,R.sub.41,R.sub.42,R.sub.44,R.sub.48,R.sub.55,R.sub.60,R.sub.66,R.sub.71=are independently C or absent; [0958] R.sub.25=C, G or absent; [0959] R.sub.9=C, G, U or absent; [0960] R.sub.8,R.sub.13,R.sub.26,R.sub.33,R.sub.35,R.sub.50,R.sub.53,R.sub.61,R.sub.68=are independently C, U or absent; [0961] R.sub.1,R.sub.6,R.sub.10,R.sub.15,R.sub.19,R.sub.20,R.sub.32,R.sub.34,R.sub.37,R.sub.39,R.sub.40,R.sub.46,R.sub.51,R.sub.52,R.sub.62,R.sub.64,R.sub.69=are independently G or absent; [0962] R.sub.16=G, U or absent; [0963] R.sub.5,R.sub.11,R.sub.21,R.sub.22,R.sub.29,R.sub.31,R.sub.36,R.sub.38,R.sub.54,R.sub.59,R.sub.65=are independently U or absent; [0964] [R.sub.47].sub.x=N or absent;

[0965] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0966] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.HIS (SEQ ID NO: 588),

[0967] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for His is: [0968] R.sub.0,R.sub.17,R.sub.18,R.sub.23=are absent [0969] R.sub.7,R.sub.12,R.sub.14,R.sub.24,R.sub.27,R.sub.45,R.sub.57,R.sub.58,R.sub.63,R.sub.67,R.sub.72=are independently A or absent; [0970] R.sub.3=A, C or absent; [0971] R.sub.4,R.sub.43,R.sub.56,R.sub.70=are independently A, G or absent; [0972] R.sub.49=A, U or absent; [0973] R.sub.2,R.sub.28,R.sub.30,R.sub.41,R.sub.42,R.sub.44,R.sub.48,R.sub.55,R.sub.60,R.sub.66,R.sub.71=are independently C or absent; [0974] R.sub.8,R.sub.9,R.sub.26,R.sub.33,R.sub.35,R.sub.50,R.sub.61,R.sub.68=are independently C, U or absent; [0975] R.sub.1,R.sub.6,R.sub.10,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.32,R.sub.34,R.sub.37,R.sub.39,R.sub.40,R.sub.46,R.sub.51,R.sub.52,R.sub.62,R.sub.64,R.sub.69=are independently G or absent; [0976] R.sub.5,R.sub.11,R.sub.13,R.sub.16,R.sub.21,R.sub.22,R.sub.29,R.sub.31,R.sub.36,R.sub.38,R.sub.53,R.sub.54,R.sub.59,R.sub.65=are independently U or absent; [0977] [R.sub.47].sub.x=N or absent;

[0978] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Isoleucine TREM Consensus Sequence

[0979] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.ILE (SEQ ID NO: 589),

[0980] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ile is: [0981] R.sub.23=absent; [0982] R.sub.38,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [0983] R.sub.1,R.sub.26=are independently A, C, G or absent; R.sub.0,R.sub.3,R.sub.4,R.sub.6,R.sub.16,R.sub.31,R.sub.32,R.sub.34,R.sub.37,R.sub.42,R.sub.43,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.55,R.sub.59,R.sub.62,R.sub.63,R.sub.64,R.sub.66,R.sub.67,R.sub.68,R.sub.69=are independently N or absent; [0984] R.sub.22,R.sub.61,R.sub.65=are independently A, C, U or absent; [0985] R.sub.9,R.sub.14,R.sub.15,R.sub.24,R.sub.27,R.sub.40=are independently A, G or absent; [0986] R.sub.7,R.sub.25,R.sub.29,R.sub.51,R.sub.56=are independently A, G, U or absent; [0987] R.sub.18,R.sub.54=are independently A, U or absent; [0988] R.sub.60=C or absent; [0989] R.sub.2,R.sub.52,R.sub.70=are independently C, G or absent; [0990] R.sub.5,R.sub.12,R.sub.21,R.sub.30,R.sub.33,R.sub.71=are independently C, G, U or absent; [0991] R.sub.11,R.sub.13,R.sub.17,R.sub.28,R.sub.35,R.sub.53,R.sub.55=are independently C, U or absent; [0992] R.sub.10,R.sub.19,R.sub.20=are independently G or absent; [0993] R.sub.8,R.sub.36,R.sub.39=are independently U or absent; [0994] [R.sub.47].sub.x=N or absent;

[0995] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[0996] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II ILE (SEQ ID NO: 590),

[0997] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ile is: [0998] R.sub.0,R.sub.18,R.sub.23=are absent [0999] R.sub.24,R.sub.38,R.sub.40,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1000] R.sub.26,R.sub.65=are independently A, C or absent; [1001] R.sub.58,R.sub.59,R.sub.67=are independently N or absent; [1002] R.sub.22=A, C, U or absent; [1003] R.sub.6,R.sub.9,R.sub.14,R.sub.15,R.sub.29,R.sub.34,R.sub.43,R.sub.46,R.sub.48,R.sub.50,R.sub.51,R.sub.63,R.sub.69=are independently A, G or absent; [1004] R.sub.37,R.sub.56=are independently A, G, U or absent; [1005] R.sub.54=A, U or absent; [1006] R.sub.28,R.sub.35,R.sub.60,R.sub.62,R.sub.71=are independently C or absent; [1007] R.sub.2,R.sub.52,R.sub.70=are independently C, G or absent; [1008] R.sub.5=C, G, U or absent; [1009] R.sub.3,R.sub.4,R.sub.11,R.sub.13,R.sub.17,R.sub.21,R.sub.30,R.sub.42,R.sub.44,R.sub.45,R.sub.49,R.sub.53,R.sub.55,R.sub.61,R.sub.64,R.sub.66=are independently C, U or absent; [1010] R.sub.1,R.sub.10,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.31,R.sub.68=are independently G or absent; [1011] R.sub.7,R.sub.12,R.sub.32=are independently G, U or absent; [1012] R.sub.8,R.sub.16,R.sub.33,R.sub.36,R.sub.39=are independently U or absent; [1013] [R.sub.47].sub.x=N or absent;

[1014] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1015] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III ILE (SEQ ID NO: 591),

[1016] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ile is: [1017] R.sub.0,R.sub.18,R.sub.23=are absent [1018] R.sub.14,R.sub.24,R.sub.38,R.sub.40,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1019] R.sub.26,R.sub.65=are independently A, C or absent; [1020] R.sub.22,R.sub.59=are independently A, C, U or absent; [1021] R.sub.6,R.sub.9,R.sub.15,R.sub.34,R.sub.43,R.sub.46,R.sub.51,R.sub.56,R.sub.63,R.sub.69=are independently A, G or absent; [1022] R.sub.37=A, G, U or absent; [1023] R.sub.13,R.sub.28,R.sub.35,R.sub.44,R.sub.55,R.sub.60,R.sub.62,R.sub.71=are independently C or absent; [1024] R.sub.2,R.sub.5,R.sub.70=are independently C, G or absent; [1025] R.sub.58,R.sub.67=are independently C, G, U or absent; [1026] R.sub.3,R.sub.4,R.sub.11,R.sub.17,R.sub.21,R.sub.30,R.sub.42,R.sub.45,R.sub.49,R.sub.53,R.sub.61,R.sub.64,R.sub.66=are independently C, U or absent; [1027] R.sub.1,R.sub.10,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.29,R.sub.31,R.sub.32,R.sub.48,R.sub.50,R.sub.52,R.sub.68=are independently G or absent; [1028] R.sub.7,R.sub.12=are independently G, U or absent; [1029] R.sub.8,R.sub.16,R.sub.33,R.sub.36,R.sub.39,R.sub.54=are independently U or absent; [1030] [R.sub.47].sub.x=N or absent;

[1031] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Methionine TREM Consensus Sequence

[1032] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.MET (SEQ ID NO: 592),

[1033] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Met is: [1034] R.sub.0,R.sub.23=are absent; [1035] R.sub.14,R.sub.38,R.sub.40,R.sub.57=are independently A or absent; [1036] R.sub.60=A, C or absent; [1037] R.sub.33,R.sub.48,R.sub.70=are independently A, C, G or absent; [1038] R.sub.1,R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.1,R.sub.12,R.sub.16,R.sub.17,R.sub.21,R.sub.22,R.sub.26,R.sub.27,R.sub.29,R.sub.3,R.sub.31,R.sub.32,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.49,R.sub.50,R.sub.58,R.sub.6 2,R.sub.63,R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.71=are independently N or absent; [1039] R.sub.18,R.sub.35,R.sub.41,R.sub.59,R.sub.65=are independently A, C, U or absent; [1040] R.sub.9,R.sub.15,R.sub.51=are independently A, G or absent; [1041] R.sub.7,R.sub.24,R.sub.25,R.sub.34,R.sub.53,R.sub.56=are independently A, G, U or absent; [1042] R.sub.72=A, U or absent; [1043] R.sub.37=C or absent; [1044] R.sub.10,R.sub.55=are independently C, G or absent; [1045] R.sub.2,R.sub.13,R.sub.28,R.sub.43,R.sub.64=are independently C, G, U or absent; [1046] R.sub.36,R.sub.61=are independently C, U or absent; [1047] R.sub.19,R.sub.20,R.sub.52=are independently G or absent; [1048] R.sub.8,R.sub.39,R.sub.54=are independently U or absent; [1049] [R.sub.47].sub.x=N or absent;

[1050] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1051] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.MET (SEQ ID NO: 593),

[1052] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Met is: [1053] R.sub.0,R.sub.18,R.sub.22,R.sub.23=are absent [1054] R.sub.14,R.sub.24,R.sub.38,R.sub.40,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1055] R.sub.59,R.sub.60,R.sub.62,R.sub.65=are independently A, C or absent; [1056] R.sub.6,R.sub.45,R.sub.67=are independently A, C, G or absent; [1057] R.sub.4=N or absent; [1058] R.sub.21,R.sub.42=are independently A, C, U or absent; [1059] R.sub.1,R.sub.9,R.sub.27,R.sub.29,R.sub.32,R.sub.46,R.sub.51=are independently A, G or absent; [1060] R.sub.17,R.sub.49,R.sub.53,R.sub.56,R.sub.58=are independently A, G, U or absent; [1061] R.sub.63=A, U or absent; [1062] R.sub.3,R.sub.13,R.sub.37=are independently C or absent; [1063] R.sub.48,R.sub.55,R.sub.64,R.sub.70=are independently C, G or absent; [1064] R.sub.2,R.sub.5,R.sub.66,R.sub.68=are independently C, G, U or absent; [1065] R.sub.11,R.sub.16,R.sub.26,R.sub.28,R.sub.30,R.sub.31,R.sub.35,R.sub.36,R.sub.43,R.sub.44,R.sub.61,R.sub.71=are independently C, U or absent; [1066] R.sub.10,R.sub.12,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.33,R.sub.52,R.sub.69=are independently G or absent; [1067] R.sub.7,R.sub.34,R.sub.50=are independently G, U or absent; [1068] R.sub.8,R.sub.39,R.sub.54=are independently U or absent; [1069] [R.sub.47].sub.x=N or absent;

[1070] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1071] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.MET (SEQ ID NO: 594),

[1072] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Met is: [1073] R.sub.0,R.sub.18,R.sub.22,R.sub.23=are absent [1074] R.sub.14,R.sub.24,R.sub.38,R.sub.40,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1075] R.sub.59,R.sub.62,R.sub.65=are independently A, C or absent; [1076] R.sub.6,R.sub.67=are independently A, C, G or absent; [1077] R.sub.4,R.sub.21=are independently A, C, U or absent; [1078] R.sub.1,R.sub.9,R.sub.27,R.sub.29,R.sub.32,R.sub.45,R.sub.46,R.sub.51=are independently A, G or absent; [1079] R.sub.17,R.sub.56,R.sub.58=are independently A, G, U or absent; [1080] R.sub.49,R.sub.53,R.sub.63=are independently A, U or absent; [1081] R.sub.3,R.sub.13,R.sub.26,R.sub.37,R.sub.43,R.sub.60=are independently C or absent; [1082] R.sub.2,R.sub.48,R.sub.55,R.sub.64,R.sub.70=are independently C, G or absent; [1083] R.sub.5,R.sub.66=are independently C, G, U or absent; [1084] R.sub.11,R.sub.16,R.sub.28,R.sub.30,R.sub.31,R.sub.35,R.sub.36,R.sub.42,R.sub.44,R.sub.61,R.sub.71=are independently C, U or absent; [1085] R.sub.10,R.sub.12,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.33,R.sub.52,R.sub.69=are independently G or absent; [1086] R.sub.7,R.sub.34,R.sub.50,R.sub.68=are independently G, U or absent; [1087] R.sub.8,R.sub.39,R.sub.54=are independently U or absent; [1088] [R.sub.47].sub.x=N or absent;

[1089] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Leucine TREM Consensus Sequence

[1090] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.LEU (SEQ ID NO: 595),

[1091] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Leu is: [1092] R.sub.0=absent; [1093] R.sub.38,R.sub.57=are independently A or absent; [1094] R.sub.60=A, C or absent; [1095] R.sub.1,R.sub.13,R.sub.27,R.sub.48,R.sub.51,R.sub.56=are independently A, C, G or absent; [1096] R.sub.2,R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.9,R.sub.10,R.sub.11,R.sub.12,R.sub.16,R.sub.23,R.sub.26,R.sub.28,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.33,R.sub.34,R.sub.37,R.sub.41,R.sub.42,R.sub.43,R.sub.44, [1097] R.sub.45,R.sub.46,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.65,R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [1098] R.sub.17,R.sub.18,R.sub.21,R.sub.22,R.sub.25,R.sub.35,R.sub.55=are independently A, C, U or absent; [1099] R.sub.14,R.sub.15,R.sub.39,R.sub.72=are independently A, G or absent; [1100] R.sub.24,R.sub.40=are independently A, G, U or absent; [1101] R.sub.52,R.sub.61,R.sub.64,R.sub.71=are independently C, G, U or absent; [1102] R.sub.36,R.sub.53,R.sub.59=are independently C, U or absent; [1103] R.sub.19=G or absent; [1104] R.sub.20=G, U or absent; [1105] R.sub.8,R.sub.54=are independently U or absent; [1106] [R.sub.47].sub.x=N or absent;

[1107] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1108] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.LEU (SEQ ID NO: 596), [1109] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72
wherein R is a ribonucleotide residue and the consensus for Leu is: [1110] R.sub.0=absent [1111] R.sub.38,R.sub.57,R.sub.72=are independently A or absent; [1112] R.sub.60=A, C or absent; [1113] R.sub.4,R.sub.5,R.sub.48,R.sub.50,R.sub.56,R.sub.69=are independently A, C, G or absent; [1114] R.sub.6,R.sub.33,R.sub.41,R.sub.43,R.sub.46,R.sub.49,R.sub.58,R.sub.63,R.sub.66,R.sub.70=are independently N or absent; [1115] R.sub.11,R.sub.12,R.sub.17,R.sub.21,R.sub.22,R.sub.28,R.sub.31,R.sub.37,R.sub.44,R.sub.55=are independently A, C, U or absent; [1116] R.sub.1,R.sub.9,R.sub.14,R.sub.15,R.sub.24,R.sub.27,R.sub.34,R.sub.39=are independently A, G or absent; [1117] R.sub.7,R.sub.29,R.sub.32,R.sub.40,R.sub.45=are independently A, G, U or absent; [1118] R.sub.25=A, U or absent; [1119] R.sub.13=C, G or absent; [1120] R.sub.2,R.sub.3,R.sub.16,R.sub.26,R.sub.30,R.sub.52,R.sub.62,R.sub.64,R.sub.65,R.sub.67,R.sub.68=are independently C, G, U or absent; [1121] R.sub.18,R.sub.35,R.sub.42,R.sub.53,R.sub.59,R.sub.61,R.sub.71=are independently C, U or absent; [1122] R.sub.19,R.sub.51=are independently G or absent; [1123] R.sub.10,R.sub.20=are independently G, U or absent; [1124] R.sub.8,R.sub.23,R.sub.36,R.sub.54=are independently U or absent; [1125] [R.sub.47].sub.x=N or absent;

[1126] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1127] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.LEU (SEQ ID NO: 597),

[1128] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Leu is: [1129] R.sub.0=absent [1130] R.sub.38,R.sub.57,R.sub.72=are independently A or absent; [1131] R.sub.60=A, C or absent; [1132] R.sub.4,R.sub.5,R.sub.48,R.sub.50,R.sub.56,R.sub.58,R.sub.69=are independently A, C, G or absent; [1133] R.sub.6,R.sub.33,R.sub.43,R.sub.46,R.sub.49,R.sub.63,R.sub.66,R.sub.70=are independently N or absent; [1134] R.sub.11,R.sub.12,R.sub.17,R.sub.21,R.sub.22,R.sub.28,R.sub.31,R.sub.37,R.sub.41,R.sub.44,R.sub.55=are independently A, C, U or absent; [1135] R.sub.1,R.sub.9,R.sub.14,R.sub.15,R.sub.24,R.sub.27,R.sub.34,R.sub.39=are independently A, G or absent; [1136] R.sub.7,R.sub.29,R.sub.32,R.sub.40,R.sub.45=are independently A, G, U or absent; [1137] R.sub.25=A, U or absent; [1138] R.sub.13=C, G or absent; [1139] R.sub.2,R.sub.3,R.sub.16,R.sub.30,R.sub.52,R.sub.62,R.sub.64,R.sub.67,R.sub.68=are independently C, G, U or absent; [1140] R.sub.18,R.sub.35,R.sub.42,R.sub.53,R.sub.59,R.sub.61,R.sub.65,R.sub.71=are independently C, U or absent; [1141] R.sub.19,R.sub.51=are independently G or absent; [1142] R.sub.10,R.sub.20,R.sub.26=are independently G, U or absent; [1143] R.sub.8,R.sub.23,R.sub.36,R.sub.54=are independently U or absent; [1144] [R.sub.47].sub.x=N or absent;

[1145] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Lysine TREM Consensus Sequence

[1146] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.LYS (SEQ ID NO: 598),

[1147] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Lys is: [1148] R.sub.0=absent [1149] R.sub.14=A or absent; [1150] R.sub.40,R.sub.41=are independently A, C or absent; [1151] R.sub.34,R.sub.43,R.sub.51=are independently A, C, G or absent; [1152] R.sub.1,R.sub.2,R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.11,R.sub.12,R.sub.16,R.sub.21,R.sub.26,R.sub.30,R.sub.31,R.sub.32,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.65, [1153] R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [1154] R.sub.13,R.sub.17,R.sub.59,R.sub.71=are independently A, C, U or absent; [1155] R.sub.9,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.52,R.sub.56=are independently A, G or absent; [1156] R.sub.24,R.sub.29,R.sub.72=are independently A, G, U or absent; [1157] R.sub.18,R.sub.57=are independently A, U or absent; [1158] R.sub.10,R.sub.33=are independently C, G or absent; [1159] R.sub.42,R.sub.61,R.sub.64=are independently C, G, U or absent; [1160] R.sub.28,R.sub.35,R.sub.36,R.sub.37,R.sub.53,R.sub.55,R.sub.60=are independently C, U or absent; [1161] R.sub.8,R.sub.22,R.sub.23,R.sub.38,R.sub.39,R.sub.54=are independently U or absent; [1162] [R.sub.47].sub.x=N or absent;

[1163] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1164] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.LYS (SEQ ID NO: 599),

[1165] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Lys is: [1166] R.sub.0,R.sub.18,R.sub.23=are absent [1167] R.sub.14=A or absent; [1168] R.sub.40,R.sub.41,R.sub.43=are independently A, C or absent; [1169] R.sub.3,R.sub.7=are independently A, C, G or absent; [1170] R.sub.1,R.sub.6,R.sub.11,R.sub.31,R.sub.45,R.sub.48,R.sub.49,R.sub.63,R.sub.65,R.sub.66,R.sub.68=are independently N or absent; [1171] R.sub.2,R.sub.12,R.sub.13,R.sub.17,R.sub.44,R.sub.67,R.sub.71=are independently A, C, U or absent; [1172] R.sub.9,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.34,R.sub.50,R.sub.52,R.sub.56,R.sub.70,R.sub.72=are independently A, G or absent; [1173] R.sub.5,R.sub.24,R.sub.26,R.sub.29,R.sub.32,R.sub.46,R.sub.69=are independently A, G, U or absent; [1174] R.sub.57=A, U or absent; [1175] R.sub.10,R.sub.61=are independently C, G or absent; [1176] R.sub.4,R.sub.16,R.sub.21,R.sub.30,R.sub.58,R.sub.64=are independently C, G, U or absent; [1177] R.sub.28,R.sub.35,R.sub.36,R.sub.37,R.sub.42,R.sub.53,R.sub.55,R.sub.59,R.sub.60,R.sub.62=are independently C, U or absent; [1178] R.sub.33,R.sub.51=are independently G or absent; [1179] R.sub.8=G, U or absent; [1180] R.sub.22,R.sub.38,R.sub.39,R.sub.54=are independently U or absent; [1181] [R.sub.47].sub.x=N or absent;

[1182] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1183] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.LYS (SEQ ID NO: 600),

[1184] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Lys is: [1185] R.sub.0,R.sub.18,R.sub.23=absent [1186] R.sub.9,R.sub.14,R.sub.34,R.sub.41=are independently A or absent; [1187] R.sub.40=A, C or absent; [1188] R.sub.1,R.sub.3,R.sub.7,R.sub.31=are independently A, C, G or absent; [1189] R.sub.48,R.sub.65,R.sub.68=are independently N or absent; [1190] R.sub.2,R.sub.13,R.sub.17,R.sub.44,R.sub.63,R.sub.66=are independently A, C, U or absent; [1191] R.sub.5,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.29,R.sub.50,R.sub.52,R.sub.56,R.sub.70,R.sub.72=are independently A, G or absent; [1192] R.sub.6,R.sub.24,R.sub.32,R.sub.49=are independently A, G, U or absent; [1193] R.sub.12,R.sub.26,R.sub.46,R.sub.57=are independently A, U or absent; [1194] R.sub.11,R.sub.28,R.sub.35,R.sub.43=are independently C or absent; [1195] R.sub.10,R.sub.45,R.sub.61=are independently C, G or absent; [1196] R.sub.4,R.sub.21,R.sub.64=are independently C, G, U or absent; [1197] R.sub.37,R.sub.53,R.sub.55,R.sub.59,R.sub.60,R.sub.62,R.sub.67,R.sub.71=are independently C, U or absent; [1198] R.sub.33,R.sub.51=are independently G or absent; [1199] R.sub.8,R.sub.30,R.sub.58,R.sub.69=are independently G, U or absent; [1200] R.sub.16,R.sub.22,R.sub.36,R.sub.38,R.sub.39,R.sub.42,R.sub.54=are independently U or absent; [1201] [R.sub.47].sub.x=N or absent;

[1202] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Phenylalanine TREM Consensus Sequence

[1203] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I PHE (SEQ ID NO: 601),

[1204] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Phe is: [1205] R.sub.0,R.sub.23=are absent [1206] R.sub.9,R.sub.14,R.sub.38,R.sub.39,R.sub.57,R.sub.72=are independently A or absent; [1207] R.sub.71=A, C or absent; [1208] R.sub.41,R.sub.70=are independently A, C, G or absent; [1209] R.sub.4,R.sub.5,R.sub.6,R.sub.30,R.sub.31,R.sub.32,R.sub.34,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.58,R.sub.62,R.sub.63,R.sub.66,R.sub.67,R.sub.68,R.sub.69=are independently N or absent; [1210] R.sub.16,R.sub.61,R.sub.65=are independently A, C, U or absent; [1211] R.sub.15,R.sub.26,R.sub.27,R.sub.29,R.sub.40,R.sub.56=are independently A, G or absent; [1212] R.sub.7,R.sub.51=are independently A, G, U or absent; [1213] R.sub.22,R.sub.24=are independently A, U or absent; [1214] R.sub.55,R.sub.60=are independently C or absent; [1215] R.sub.2,R.sub.3,R.sub.21,R.sub.33,R.sub.43,R.sub.50,R.sub.64=are independently C, G, U or absent; [1216] R.sub.11,R.sub.12,R.sub.13,R.sub.17,R.sub.28,R.sub.35,R.sub.36,R.sub.59=are independently C, U or absent; [1217] R.sub.10,R.sub.19,R.sub.20,R.sub.25,R.sub.37,R.sub.52=are independently G or absent; [1218] R.sub.1=G, U or absent; [1219] R.sub.8,R.sub.18,R.sub.53,R.sub.54=are independently U or absent; [1220] [R.sub.47].sub.x=N or absent;

[1221] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1222] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.PHE (SEQ ID NO: 602),

[1223] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Phe is: [1224] R.sub.0,R.sub.18,R.sub.23=absent [1225] R.sub.14,R.sub.24,R.sub.38,R.sub.39,R.sub.57,R.sub.72=are independently A or absent; [1226] R.sub.46,R.sub.71=are independently A, C or absent; [1227] R.sub.4,R.sub.70=are independently A, C, G or absent; [1228] R.sub.45=A, C, U or absent; [1229] R.sub.6,R.sub.7,R.sub.15,R.sub.26,R.sub.27,R.sub.32,R.sub.34,R.sub.40,R.sub.41,R.sub.56,R.sub.69=are independently A, G or absent; [1230] R.sub.29=A, G, U or absent; [1231] R.sub.5,R.sub.9,R.sub.67=are independently A, U or absent; [1232] R.sub.35,R.sub.49,R.sub.55,R.sub.60=are independently C or absent; [1233] R.sub.21,R.sub.43,R.sub.62=are independently C, G or absent; [1234] R.sub.2,R.sub.33,R.sub.68=are independently C, G, U or absent; [1235] R.sub.3,R.sub.11,R.sub.12,R.sub.13,R.sub.28,R.sub.30,R.sub.36,R.sub.42,R.sub.44,R.sub.48,R.sub.58,R.sub.59,R.sub.61,R.sub.66=are independently C, U or absent; [1236] R.sub.10,R.sub.19,R.sub.20,R.sub.25,R.sub.37,R.sub.51,R.sub.52,R.sub.63,R.sub.64=are independently G or absent; [1237] R.sub.1,R.sub.31,R.sub.50=are independently G, U or absent; [1238] R.sub.8,R.sub.16,R.sub.17,R.sub.22,R.sub.53,R.sub.54,R.sub.65=are independently U or absent; [1239] [R.sub.47].sub.x=N or absent;

[1240] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1241] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III PHE (SEQ ID NO: 603),

[1242] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Phe is: [1243] R.sub.0,R.sub.18,R.sub.22,R.sub.23=absent [1244] R.sub.5,R.sub.7,R.sub.14,R.sub.24,R.sub.26,R.sub.32,R.sub.34,R.sub.38,R.sub.39,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1245] R.sub.46=A, C or absent; [1246] R.sub.70=A, C, G or absent; [1247] R.sub.4,R.sub.6,R.sub.15,R.sub.56,R.sub.69=are independently A, G or absent; [1248] R.sub.9,R.sub.45=are independently A, U or absent; [1249] R.sub.2,R.sub.11,R.sub.13,R.sub.35,R.sub.43,R.sub.49,R.sub.55,R.sub.60,R.sub.68,R.sub.71=are independently C or absent; [1250] R.sub.33=C, G or absent; [1251] R.sub.3,R.sub.28,R.sub.36,R.sub.48,R.sub.58,R.sub.59,R.sub.61=are independently C, U or absent; [1252] R.sub.1,R.sub.10,R.sub.19,R.sub.20,R.sub.21,R.sub.25,R.sub.27,R.sub.29,R.sub.37,R.sub.40,R.sub.51,R.sub.52,R.sub.62,R.sub.63,R.sub.64=are independently G or absent; [1253] R.sub.8,R.sub.12,R.sub.16,R.sub.17,R.sub.30,R.sub.31,R.sub.42,R.sub.44,R.sub.50,R.sub.53,R.sub.54,R.sub.65,R.sub.66,R.sub.67=are independently U or absent; [1254] [R.sub.47].sub.x=N or absent;

[1255] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Proline TREM Consensus Sequence

[1256] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.PRO (SEQ ID NO: 604),

[1257] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43- R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Pro is: [1258] R.sub.0=absent [1259] R.sub.14,R.sub.57=are independently A or absent; [1260] R.sub.70,R.sub.72=are independently A, C or absent; [1261] R.sub.9,R.sub.26,R.sub.27=are independently A, C, G or absent; [1262] R.sub.4,R.sub.5,R.sub.6,R.sub.16,R.sub.21,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.33,R.sub.34,R.sub.37,R.sub.41,R.sub.42,R.sub.43,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.58,R.sub.61,R.sub.62, [1263] R.sub.63,R.sub.64,R.sub.66,R.sub.67,R.sub.68=are independently N or absent; [1264] R.sub.35,R.sub.65=are independently A, C, U or absent; [1265] R.sub.24,R.sub.40,R.sub.56=are independently A, G or absent; [1266] R.sub.7,R.sub.25,R.sub.51=are independently A, G, U or absent; [1267] R.sub.55,R.sub.60=are independently C or absent; [1268] R.sub.1,R.sub.3,R.sub.71=are independently C, G or absent; [1269] R.sub.11,R.sub.12,R.sub.20,R.sub.69=are independently C, G, U or absent; [1270] R.sub.13,R.sub.17,R.sub.18,R.sub.22,R.sub.23,R.sub.28,R.sub.59=are independently C, U or absent; [1271] R.sub.10,R.sub.15,R.sub.19,R.sub.38,R.sub.39,R.sub.52=are independently G or absent; [1272] R.sub.2=are independently G, U or absent; [1273] R.sub.8,R.sub.36,R.sub.53,R.sub.54=are independently U or absent; [1274] [R.sub.47].sub.x=N or absent;

[1275] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1276] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.PRO (SEQ ID NO: 605),

[1277] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Pro is: [1278] R.sub.0,R.sub.17,R.sub.18,R.sub.22,R.sub.23=absent; [1279] R.sub.14,R.sub.45,R.sub.56,R.sub.57,R.sub.58,R.sub.65,R.sub.68=are independently A or absent; [1280] R.sub.61=A, C, G or absent; [1281] R.sub.43=N or absent; [1282] R.sub.37=A, C, U or absent; [1283] R.sub.24,R.sub.27,R.sub.33,R.sub.40,R.sub.44,R.sub.63=are independently A, G or absent; [1284] R.sub.3,R.sub.12,R.sub.30,R.sub.32,R.sub.48,R.sub.55,R.sub.60,R.sub.70,R.sub.71,R.sub.72=are independently C or absent; [1285] R.sub.5,R.sub.34,R.sub.42,R.sub.66=are independently C, G or absent; [1286] R.sub.20=C, G, U or absent; [1287] R.sub.35,R.sub.41,R.sub.49,R.sub.62=are independently C, U or absent; [1288] R.sub.1,R.sub.2,R.sub.6,R.sub.9,R.sub.10,R.sub.15,R.sub.19,R.sub.26,R.sub.38,R.sub.39,R.sub.46,R.sub.50,R.sub.51,R.sub.52,R.sub.64,R.sub.67,R.sub.69=are independently G or absent; [1289] R.sub.11,R.sub.16=are independently G, U or absent; [1290] R.sub.4,R.sub.7,R.sub.8,R.sub.13,R.sub.21,R.sub.25,R.sub.28,R.sub.29,R.sub.31,R.sub.36,R.sub.53,R.sub.54,R.sub.59=are independently U or absent; [1291] [R.sub.47].sub.x=N or absent;

[1292] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1293] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.PRO (SEQ ID NO: 606),

[1294] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Pro is: [1295] R.sub.0,R.sub.17,R.sub.18,R.sub.22,R.sub.23=absent [1296] R.sub.14,R.sub.45,R.sub.56,R.sub.57,R.sub.58,R.sub.65,R.sub.68=are independently A or absent; [1297] R.sub.37=A, C, U or absent; [1298] R.sub.24,R.sub.27,R.sub.40=are independently A, G or absent; [1299] R.sub.3,R.sub.5,R.sub.12,R.sub.30,R.sub.32,R.sub.48,R.sub.49,R.sub.55,R.sub.60,R.sub.61,R.sub.62,R.sub.66,R.sub.70,R.sub.71,R.sub.72=are independently C or absent; [1300] R.sub.34,R.sub.42=are independently C, G or absent; [1301] R.sub.43=C, G, U or absent; [1302] R.sub.41=C, U or absent; [1303] R.sub.1,R.sub.2,R.sub.6,R.sub.9,R.sub.10,R.sub.15,R.sub.19,R.sub.20,R.sub.26,R.sub.33,R.sub.38,R.sub.39,R.sub.44,R.sub.46,R.sub.50,R.sub.51,R.sub.52,R.sub.63,R.sub.64,R.sub.67,R.sub.69=are independently G or absent; [1304] R.sub.16=G, U or absent; [1305] R.sub.4,R.sub.7,R.sub.8,R.sub.11,R.sub.13,R.sub.21,R.sub.25,R.sub.28,R.sub.29,R.sub.31,R.sub.35,R.sub.36,R.sub.53,R.sub.54,R.sub.59=are independently U or absent; [1306] [R.sub.47].sub.x=N or absent;

[1307] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Serine TREM Consensus Sequence

[1308] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.SER (SEQ ID NO: 607),

[1309] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ser is: [1310] R.sub.0=absent; [1311] R.sub.14,R.sub.24,R.sub.57=are independently A or absent; [1312] R.sub.41=A, C or absent; [1313] R.sub.2,R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.9,R.sub.10,R.sub.11,R.sub.12,R.sub.13,R.sub.16,R.sub.21,R.sub.25,R.sub.26,R.sub.27,R.sub.28,R.sub.3,R.sub.31,R.sub.32,R.sub.33,R.sub.34,R.sub.37,R.sub.42,R.sub.43, R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.62,R.sub.63,R.sub.64,R.sub.65,R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [1314] R.sub.18=A, C, U or absent; [1315] R.sub.15,R.sub.40,R.sub.51,R.sub.56=are independently A, G or absent; [1316] R.sub.1,R.sub.29,R.sub.58,R.sub.72=are independently A, G, U or absent; [1317] R.sub.39=A, U or absent; [1318] R.sub.60=C or absent; [1319] R.sub.38=C, G or absent; [1320] R.sub.17,R.sub.22,R.sub.23,R.sub.71=are independently C, G, U or absent; [1321] R.sub.8,R.sub.35,R.sub.36,R.sub.55,R.sub.59,R.sub.61=are independently C, U or absent; [1322] R.sub.19,R.sub.20=are independently G or absent; [1323] R.sub.52=G, U or absent; [1324] R.sub.53,R.sub.54=are independently U or absent; [1325] [R.sub.47].sub.x=N or absent;

[1326] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1327] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.SER (SEQ ID NO: 608),

[1328] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ser is: [1329] R.sub.0,R.sub.23=absent [1330] R.sub.14,R.sub.24,R.sub.41,R.sub.57=are independently A or absent; [1331] R.sub.44=A, C or absent; [1332] R.sub.25,R.sub.45,R.sub.48=are independently A, C, G or absent; [1333] R.sub.2,R.sub.3,R.sub.4,R.sub.5,R.sub.37,R.sub.50,R.sub.62,R.sub.66,R.sub.67,R.sub.69,R.sub.70=are independently N or absent; [1334] R.sub.12,R.sub.28,R.sub.65=are independently A, C, U or absent; [1335] R.sub.9,R.sub.15,R.sub.29,R.sub.34,R.sub.40,R.sub.56,R.sub.63=are independently A, G or absent; [1336] R.sub.7,R.sub.26,R.sub.30,R.sub.33,R.sub.46,R.sub.58,R.sub.72=are independently A, G, U or absent; [1337] R.sub.39=A, U or absent; [1338] R.sub.11,R.sub.35,R.sub.60,R.sub.61=are independently C or absent; [1339] R.sub.13,R.sub.38=are independently C, G or absent; [1340] R.sub.6,R.sub.17,R.sub.31,R.sub.43,R.sub.64,R.sub.68=are independently C, G, U or absent; [1341] R.sub.36,R.sub.42,R.sub.49,R.sub.55,R.sub.59,R.sub.71=are independently C, U or absent; [1342] R.sub.10,R.sub.19,R.sub.20,R.sub.27,R.sub.51=are independently G or absent; [1343] R.sub.1,R.sub.16,R.sub.32,R.sub.52=are independently G, U or absent; [1344] R.sub.8,R.sub.18,R.sub.21,R.sub.22,R.sub.53,R.sub.54=are independently U or absent; [1345] [R.sub.47].sub.x=N or absent;

[1346] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1347] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.SER (SEQ ID NO: 609),

[1348] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Ser is: [1349] R.sub.0,R.sub.23=absent [1350] R.sub.14,R.sub.24,R.sub.41,R.sub.57,R.sub.58=are independently A or absent; [1351] R.sub.44=A, C or absent; [1352] R.sub.25,R.sub.48=are independently A, C, G or absent; [1353] R.sub.2,R.sub.3,R.sub.5,R.sub.37,R.sub.66,R.sub.67,R.sub.69,R.sub.70=are independently N or absent; [1354] R.sub.12,R.sub.28,R.sub.62=are independently A, C, U or absent; [1355] R.sub.7,R.sub.9,R.sub.15,R.sub.29,R.sub.33,R.sub.34,R.sub.40,R.sub.45,R.sub.56,R.sub.63=are independently A, G or absent; [1356] R.sub.4,R.sub.26,R.sub.46,R.sub.50=are independently A, G, U or absent; [1357] R.sub.30,R.sub.39=are independently A, U or absent; [1358] R.sub.11,R.sub.17,R.sub.35,R.sub.60,R.sub.61=are independently C or absent; [1359] R.sub.13,R.sub.38=are independently C, G or absent; [1360] R.sub.6,R.sub.64=are independently C, G, U or absent; [1361] R.sub.31,R.sub.42,R.sub.43,R.sub.49,R.sub.55,R.sub.59,R.sub.65,R.sub.68,R.sub.71=are independently C, U or absent; [1362] R.sub.10,R.sub.19,R.sub.20,R.sub.27,R.sub.51,R.sub.52=are independently G or absent; [1363] R.sub.1,R.sub.16,R.sub.32,R.sub.72=are independently G, U or absent; [1364] R.sub.8,R.sub.18,R.sub.21,R.sub.22,R.sub.36,R.sub.53,R.sub.54=are independently U or absent; [1365] [R.sub.47].sub.x=N or absent;

[1366] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Threonine TREM Consensus Sequence

[1367] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.THR (SEQ ID NO: 610),

[1368] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Thr is: [1369] R.sub.0,R.sub.23=absent [1370] R.sub.14,R.sub.41,R.sub.57=are independently A or absent; [1371] R.sub.56,R.sub.70=are independently A, C, G or absent; [1372] R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.12,R.sub.16,R.sub.26,R.sub.30,R.sub.31,R.sub.32,R.sub.34,R.sub.37,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.64,R.sub.65,R.sub.66,R.sub.67,R.sub.68,R.sub.72=are independently N or absent; [1373] R.sub.13,R.sub.17,R.sub.21,R.sub.35,R.sub.61=are independently A, C, U or absent; [1374] R.sub.1,R.sub.9,R.sub.24,R.sub.27,R.sub.29,R.sub.69=are independently A, G or absent; [1375] R.sub.15,R.sub.25,R.sub.51=are independently A, G, U or absent; [1376] R.sub.40,R.sub.53=are independently A, U or absent; [1377] R.sub.33,R.sub.43=are independently C, G or absent; [1378] R.sub.2,R.sub.3,R.sub.59=are independently C, G, U or absent; [1379] R.sub.11,R.sub.18,R.sub.22,R.sub.28,R.sub.36,R.sub.54,R.sub.55,R.sub.60,R.sub.71=are independently C, U or absent; [1380] R.sub.10,R.sub.20,R.sub.38,R.sub.52=are independently G or absent; [1381] R.sub.19=G, U or absent; [1382] R.sub.8,R.sub.39=are independently U or absent; [1383] [R.sub.47].sub.x=N or absent;

[1384] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1385] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II THR (SEQ ID NO: 611),

[1386] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Thr is: [1387] R.sub.0,R.sub.18,R.sub.23=absent [1388] R.sub.14,R.sub.41,R.sub.57=are independently A or absent; [1389] R.sub.9,R.sub.42,R.sub.44,R.sub.48,R.sub.56,R.sub.70=are independently A, C, G or absent; [1390] R.sub.4,R.sub.6,R.sub.12,R.sub.26,R.sub.49,R.sub.58,R.sub.63,R.sub.64,R.sub.66,R.sub.68=are independently N or absent; [1391] R.sub.13,R.sub.21,R.sub.31,R.sub.37,R.sub.62=are independently A, C, U or absent; [1392] R.sub.1,R.sub.15,R.sub.24,R.sub.27,R.sub.29,R.sub.46,R.sub.51,R.sub.69=are independently A, G or absent; [1393] R.sub.7,R.sub.25,R.sub.45,R.sub.50,R.sub.67=are independently A, G, U or absent; [1394] R.sub.40,R.sub.53=are independently A, U or absent; [1395] R.sub.35=C or absent; [1396] R.sub.33,R.sub.43=are independently C, G or absent; [1397] R.sub.2,R.sub.3,R.sub.5,R.sub.16,R.sub.32,R.sub.34,R.sub.59,R.sub.65,R.sub.72=are independently C, G, U or absent; [1398] R.sub.11,R.sub.17,R.sub.22,R.sub.28,R.sub.30,R.sub.36,R.sub.55,R.sub.60,R.sub.61,R.sub.71=are independently C, U or absent; [1399] R.sub.10,R.sub.19,R.sub.20,R.sub.38,R.sub.52=are independently G or absent; [1400] R.sub.8,R.sub.39,R.sub.54=are independently U or absent; [1401] [R.sub.47].sub.x=N or absent;

[1402] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1403] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.THR (SEQ ID NO: 612),

[1404] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Thr is: [1405] R.sub.0,R.sub.18,R.sub.23=absent [1406] R.sub.14,R.sub.40,R.sub.41,R.sub.57=are independently A or absent; [1407] R.sub.44=A, C or absent; [1408] R.sub.9,R.sub.42,R.sub.48,R.sub.56=are independently A, C, G or absent; [1409] R.sub.4,R.sub.6,R.sub.12,R.sub.26,R.sub.58,R.sub.64,R.sub.66,R.sub.68=are independently N or absent; [1410] R.sub.13,R.sub.21,R.sub.31,R.sub.37,R.sub.49,R.sub.62=are independently A, C, U or absent; [1411] R.sub.1,R.sub.15,R.sub.24,R.sub.27,R.sub.29,R.sub.46,R.sub.51,R.sub.69=are independently A, G or absent; [1412] R.sub.7,R.sub.25,R.sub.45,R.sub.50,R.sub.63,R.sub.67=are independently A, G, U or absent; [1413] R.sub.53=A, U or absent; [1414] R.sub.35=C or absent; [1415] R.sub.2,R.sub.33,R.sub.43,R.sub.70=are independently C, G or absent; [1416] R.sub.5,R.sub.16,R.sub.34,R.sub.59,R.sub.65=are independently C, G, U or absent; [1417] R.sub.3,R.sub.11,R.sub.22,R.sub.28,R.sub.30,R.sub.36,R.sub.55,R.sub.60,R.sub.61,R.sub.71=are independently C, U or absent; [1418] R.sub.10,R.sub.19,R.sub.20,R.sub.38,R.sub.52=are independently G or absent; [1419] R.sub.32=G, U or absent; [1420] R.sub.8,R.sub.17,R.sub.39,R.sub.54,R.sub.72=are independently U or absent; [1421] [R.sub.47].sub.x=N or absent;

[1422] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Tryptophan TREM Consensus Sequence

[1423] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.TRP (SEQ ID NO: 613),

[1424] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.1-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Trp is: [1425] R.sub.0=absent; [1426] R.sub.24,R.sub.39,R.sub.41,R.sub.57=are independently A or absent; [1427] R.sub.2,R.sub.3,R.sub.26,R.sub.27,R.sub.40,R.sub.48=are independently A, C, G or absent; [1428] R.sub.4,R.sub.5,R.sub.6,R.sub.29,R.sub.30,R.sub.31,R.sub.32,R.sub.34,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.49,R.sub.51,R.sub.58,R.sub.63,R.sub.66,R.sub.67,R.sub.68=are independently N or absent; [1429] R.sub.13,R.sub.14,R.sub.16,R.sub.18,R.sub.21,R.sub.61,R.sub.65,R.sub.71=are independently A, C, U or absent; [1430] R.sub.1,R.sub.9,R.sub.10,R.sub.15,R.sub.33,R.sub.50,R.sub.56=are independently A, G or absent; [1431] R.sub.7,R.sub.25,R.sub.72=are independently A, G, U or absent; [1432] R.sub.37,R.sub.38,R.sub.55,R.sub.60=are independently C or absent; [1433] R.sub.12,R.sub.35,R.sub.43,R.sub.64,R.sub.69,R.sub.70=are independently C, G, U or absent; [1434] R.sub.11,R.sub.17,R.sub.22,R.sub.28,R.sub.59,R.sub.62=are independently C, U or absent; [1435] R.sub.19,R.sub.20,R.sub.52=are independently G or absent; [1436] R.sub.8,R.sub.23,R.sub.36,R.sub.53,R.sub.54=are independently U or absent; [1437] [R.sub.47].sub.x=N or absent;

[1438] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1439] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II TRP (SEQ ID NO: 614),

[1440] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Trp is: [1441] R.sub.0,R.sub.18,R.sub.22,R.sub.23=absent [1442] R.sub.14,R.sub.24,R.sub.39,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1443] R.sub.3,R.sub.4,R.sub.13,R.sub.61,R.sub.71=are independently A, C or absent; [1444] R.sub.6,R.sub.44=are independently A, C, G or absent; [1445] R.sub.21=A, C, U or absent; [1446] R.sub.2,R.sub.7,R.sub.15,R.sub.25,R.sub.33,R.sub.34,R.sub.45,R.sub.56,R.sub.63=are independently A, G or absent; [1447] R.sub.58=A, G, U or absent; [1448] R.sub.46=A, U or absent; [1449] R.sub.37,R.sub.38,R.sub.55,R.sub.60,R.sub.62=are independently C or absent; [1450] R.sub.12,R.sub.26,R.sub.27,R.sub.35,R.sub.40,R.sub.48,R.sub.67=are independently C, G or absent; [1451] R.sub.32,R.sub.43,R.sub.68=are independently C, G, U or absent; [1452] R.sub.11,R.sub.16,R.sub.28,R.sub.31,R.sub.49,R.sub.59,R.sub.65,R.sub.70=are independently C, U or absent; [1453] R.sub.1,R.sub.9,R.sub.10,R.sub.19,R.sub.20,R.sub.50,R.sub.52,R.sub.69=are independently G or absent; [1454] R.sub.5,R.sub.8,R.sub.29,R.sub.30,R.sub.42,R.sub.51,R.sub.64,R.sub.66=are independently G, U or absent; [1455] R.sub.17,R.sub.36,R.sub.53,R.sub.54=are independently U or absent; [1456] [R.sub.47].sub.x=N or absent;

[1457] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1458] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.TRP (SEQ ID NO: 615),

[1459] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Trp is: [1460] R.sub.0,R.sub.18,R.sub.22,R.sub.23=absent [1461] R.sub.14,R.sub.24,R.sub.39,R.sub.41,R.sub.57,R.sub.72=are independently A or absent; [1462] R.sub.3,R.sub.4,R.sub.13,R.sub.61,R.sub.71=are independently A, C or absent; [1463] R.sub.6,R.sub.44=are independently A, C, G or absent; [1464] R.sub.21=A, C, U or absent; [1465] R.sub.2,R.sub.7,R.sub.15,R.sub.25,R.sub.33,R.sub.34,R.sub.45,R.sub.56,R.sub.63=are independently A, G or absent; [1466] R.sub.58=A, G, U or absent; [1467] R.sub.46=A, U or absent; [1468] R.sub.37,R.sub.38,R.sub.55,R.sub.60,R.sub.62=are independently C or absent; [1469] R.sub.12,R.sub.26,R.sub.27,R.sub.35,R.sub.40,R.sub.48,R.sub.67=are independently C, G or absent; [1470] R.sub.32,R.sub.43,R.sub.68=are independently C, G, U or absent; [1471] R.sub.11,R.sub.16,R.sub.28,R.sub.31,R.sub.49,R.sub.59,R.sub.65,R.sub.70=are independently C, U or absent; [1472] R.sub.1,R.sub.9,R.sub.10,R.sub.19,R.sub.20,R.sub.50,R.sub.52,R.sub.69=are independently G or absent; [1473] R.sub.5,R.sub.8,R.sub.29,R.sub.30,R.sub.42,R.sub.51,R.sub.64,R.sub.66=are independently G, U or absent; [1474] R.sub.17,R.sub.36,R.sub.53,R.sub.54=are independently U or absent; [1475] [R.sub.47].sub.x=N or absent;

[1476] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Tyrosine TREM Consensus Sequence

[1477] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.TYR (SEQ ID NO: 616),

[1478] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Tyr is: [1479] R.sub.0=absent [1480] R.sub.14,R.sub.39,R.sub.57=are independently A or absent; [1481] R.sub.41,R.sub.48,R.sub.51,R.sub.71=are independently A, C, G or absent; [1482] R.sub.3,R.sub.4,R.sub.5,R.sub.6,R.sub.9,R.sub.10,R.sub.12,R.sub.13,R.sub.16,R.sub.25,R.sub.26,R.sub.3,R.sub.31,R.sub.32,R.sub.42,R.sub.44,R.sub.45,R.sub.46,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.63,R.sub.66, [1483] R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [1484] R.sub.22,R.sub.65=are independently A, C, U or absent; [1485] R.sub.15,R.sub.24,R.sub.27,R.sub.33,R.sub.37,R.sub.40,R.sub.56=are independently A, G or absent; [1486] R.sub.7,R.sub.29,R.sub.34,R.sub.72=are independently A, G, U or absent; [1487] R.sub.23,R.sub.53=are independently A, U or absent; [1488] R.sub.35,R.sub.60=are independently C or absent; [1489] R.sub.20=C, G or absent; [1490] R.sub.1,R.sub.2,R.sub.28,R.sub.61,R.sub.64=are independently C, G, U or absent; [1491] R.sub.11,R.sub.17,R.sub.21,R.sub.43,R.sub.55=are independently C, U or absent; [1492] R.sub.19,R.sub.52=are independently G or absent; [1493] R.sub.8,R.sub.18,R.sub.36,R.sub.38,R.sub.54,R.sub.59=are independently U or absent; [1494] [R.sub.47].sub.x=N or absent;

[1495] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1496] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.TYR (SEQ ID NO: 617),

[1497] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Tyr is: [1498] R.sub.0,R.sub.18,R.sub.23=absent [1499] R.sub.7,R.sub.9,R.sub.14,R.sub.24,R.sub.26,R.sub.34,R.sub.39,R.sub.57=are independently A or absent; [1500] R.sub.44,R.sub.69=are independently A, C or absent; [1501] R.sub.71=A, C, G or absent; [1502] R.sub.68=N or absent; [1503] R.sub.58=A, C, U or absent; [1504] R.sub.33,R.sub.37,R.sub.41,R.sub.56,R.sub.62,R.sub.63=are independently A, G or absent; [1505] R.sub.6,R.sub.29,R.sub.72=are independently A, G, U or absent; [1506] R.sub.31,R.sub.45,R.sub.53=are independently A, U or absent; [1507] R.sub.13,R.sub.35,R.sub.49,R.sub.60=are independently C or absent; [1508] R.sub.20,R.sub.48,R.sub.64,R.sub.67,R.sub.70=are independently C, G or absent; [1509] R.sub.1,R.sub.2,R.sub.5,R.sub.16,R.sub.66=are independently C, G, U or absent; [1510] R.sub.11,R.sub.21,R.sub.28,R.sub.43,R.sub.55,R.sub.61=are independently C, U or absent; [1511] R.sub.10,R.sub.15,R.sub.19,R.sub.25,R.sub.27,R.sub.40,R.sub.51,R.sub.52=are independently G or absent; [1512] R.sub.3,R.sub.4,R.sub.30,R.sub.32,R.sub.42,R.sub.46=are independently G, U or absent; [1513] R.sub.8,R.sub.12,R.sub.17,R.sub.22,R.sub.36,R.sub.38,R.sub.50,R.sub.54,R.sub.59,R.sub.65=are independently U or absent; [1514] [R.sub.47],x=N or absent;

[1515] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1516] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.TYR (SEQ ID NO: 618),

[1517] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Tyr is: [1518] R.sub.0,R.sub.18,R.sub.23=absent [1519] R.sub.7,R.sub.9,R.sub.14,R.sub.24,R.sub.26,R.sub.34,R.sub.39,R.sub.57,R.sub.72=are independently A or absent; [1520] R.sub.44,R.sub.69=are independently A, C or absent; [1521] R.sub.71=A, C, G or absent; [1522] R.sub.37,R.sub.41,R.sub.56,R.sub.62,R.sub.63=are independently A, G or absent; [1523] R.sub.6,R.sub.29,R.sub.68=are independently A, G, U or absent; [1524] R.sub.31,R.sub.45,R.sub.58=are independently A, U or absent; [1525] R.sub.13,R.sub.28,R.sub.35,R.sub.49,R.sub.60,R.sub.61=are independently C or absent; [1526] R.sub.5,R.sub.48,R.sub.64,R.sub.67,R.sub.70=are independently C, G or absent; [1527] R.sub.1,R.sub.2=are independently C, G, U or absent; [1528] R.sub.11,R.sub.16,R.sub.21,R.sub.43,R.sub.55,R.sub.66=are independently C, U or absent; [1529] R.sub.10,R.sub.15,R.sub.19,R.sub.20,R.sub.25,R.sub.27,R.sub.33,R.sub.40,R.sub.51,R.sub.52=are independently G or absent; [1530] R.sub.3,R.sub.4,R.sub.30,R.sub.32,R.sub.42,R.sub.46=are independently G, U or absent; [1531] R.sub.8,R.sub.12,R.sub.17,R.sub.22,R.sub.36,R.sub.38,R.sub.50,R.sub.53,R.sub.54,R.sub.59,R.sub.65=are independently U or absent; [1532] [R.sub.47].sub.x=N or absent;

[1533] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Valine TREM Consensus Sequence

[1534] In an embodiment, a TREM disclosed herein comprises the sequence of Formula I.sub.VAL (SEQ ID NO: 619),

[1535] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Val is: [1536] R.sub.0,R.sub.23=absent; [1537] R.sub.24,R.sub.38,R.sub.57=are independently A or absent; [1538] R.sub.9,R.sub.72=are independently A, C, G or absent; [1539] R.sub.2,R.sub.4,R.sub.5,R.sub.6,R.sub.7,R.sub.12,R.sub.15,R.sub.16,R.sub.21,R.sub.25,R.sub.26,R.sub.29,R.sub.31,R.sub.32,R.sub.33,R.sub.34,R.sub.37,R.sub.41,R.sub.42,R.sub.43,R.sub.44,R.sub.45,R.sub.46,R.sub.48,R.sub.49,R.sub.50,R.sub.58,R.sub.61,R.sub.62,R.sub.63,R.sub.64,R.sub.65,R.sub.66,R.sub.67,R.sub.68,R.sub.69,R.sub.70=are independently N or absent; [1540] R.sub.17,R.sub.35,R.sub.59=are independently A, C, U or absent; [1541] R.sub.10,R.sub.14,R.sub.27,R.sub.40,R.sub.52,R.sub.56=are independently A, G or absent; [1542] R.sub.1,R.sub.3,R.sub.51,R.sub.53=are independently A, G, U or absent; [1543] R.sub.39=C or absent; [1544] R.sub.13,R.sub.30,R.sub.55=are independently C, G, U or absent; [1545] R.sub.11,R.sub.22,R.sub.28,R.sub.60,R.sub.71=are independently C, U or absent; [1546] R.sub.19=G or absent; [1547] R.sub.20=G, U or absent; [1548] R.sub.8,R.sub.18,R.sub.36,R.sub.54=are independently U or absent; [1549] [R.sub.47].sub.x=N or absent;

[1550] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1551] In an embodiment, a TREM disclosed herein comprises the sequence of Formula II.sub.VAL (SEQ ID NO: 620),

[1552] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Val is: [1553] R.sub.0,R.sub.18,R.sub.23=absent; [1554] R.sub.24,R.sub.38,R.sub.57=are independently A or absent; [1555] R.sub.64,R.sub.70,R.sub.72=are independently A, C, G or absent; [1556] R.sub.15,R.sub.16,R.sub.26,R.sub.29,R.sub.31,R.sub.32,R.sub.43,R.sub.44,R.sub.45,R.sub.49,R.sub.50,R.sub.58,R.sub.62,R.sub.65=are independently N or absent; [1557] R.sub.6,R.sub.17,R.sub.34,R.sub.37,R.sub.41,R.sub.59=are independently A, C, U or absent; [1558] R.sub.9,R.sub.10,R.sub.14,R.sub.27,R.sub.40,R.sub.46,R.sub.51,R.sub.52,R.sub.56=are independently A, G or absent; [1559] R.sub.7,R.sub.12,R.sub.25,R.sub.33,R.sub.53,R.sub.63,R.sub.66,R.sub.68=are independently A, G, U or absent; [1560] R.sub.69=A, U or absent; [1561] R.sub.39=C or absent; [1562] R.sub.5,R.sub.67=are independently C, G or absent; [1563] R.sub.2,R.sub.4,R.sub.13,R.sub.48,R.sub.55,R.sub.61=are independently C, G, U or absent; [1564] R.sub.11,R.sub.22,R.sub.28,R.sub.30,R.sub.35,R.sub.60,R.sub.71=are independently C, U or absent; [1565] R.sub.19=G or absent; [1566] R.sub.1,R.sub.3,R.sub.20,R.sub.42=are independently G, U or absent; [1567] R.sub.8,R.sub.21,R.sub.36,R.sub.54=are independently U or absent; [1568] [R.sub.47].sub.x=N or absent;

[1569] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

[1570] In an embodiment, a TREM disclosed herein comprises the sequence of Formula III.sub.VAL (SEQ ID NO: 621),

[1571] R.sub.0-R.sub.1-R.sub.2-R.sub.3-R.sub.4-R.sub.5-R.sub.6-R.sub.7-R.sub.8-R.sub.9-R.sub.10-R.sub.11-R.sub.12-R.sub.13-R.sub.14-R.sub.15-R.sub.16-R.sub.17-R.sub.18-R.sub.19-R.sub.20-R.sub.21-R.sub.22-R.sub.23-R.sub.24-R.sub.25-R.sub.26-R.sub.27-R.sub.28-R.sub.29-R.sub.30-R.sub.31-R.sub.32-R.sub.33-R.sub.34-R.sub.35-R.sub.36-R.sub.37-R.sub.38-R.sub.39-R.sub.40-R.sub.41-R.sub.42-R.sub.43-R.sub.44-R.sub.45-R.sub.46-[R.sub.47].sub.x-R.sub.48-R.sub.49-R.sub.50-R.sub.51-R.sub.52-R.sub.53-R.sub.54-R.sub.55-R.sub.56-R.sub.57-R.sub.58-R.sub.59-R.sub.60-R.sub.61-R.sub.62-R.sub.63-R.sub.64-R.sub.65-R.sub.66-R.sub.67-R.sub.68-R.sub.69-R.sub.70-R.sub.71-R.sub.72

wherein R is a ribonucleotide residue and the consensus for Val is: [1572] R.sub.0,R.sub.18,R.sub.23=absent [1573] R.sub.24,R.sub.38,R.sub.40,R.sub.57,R.sub.72=are independently A or absent; [1574] R.sub.29,R.sub.64,R.sub.70=are independently A, C, G or absent; [1575] R.sub.49,R.sub.50,R.sub.62=are independently N or absent; [1576] R.sub.16,R.sub.26,R.sub.31,R.sub.32,R.sub.37,R.sub.41,R.sub.43,R.sub.59,R.sub.65=are independently A, C, U or absent; [1577] R.sub.9,R.sub.14,R.sub.27,R.sub.46,R.sub.52,R.sub.56,R.sub.66=are independently A, G or absent; [1578] R.sub.7,R.sub.12,R.sub.25,R.sub.33,R.sub.44,R.sub.45,R.sub.53,R.sub.58,R.sub.63,R.sub.68=are independently A, G, U or absent; [1579] R.sub.69=A, U or absent; [1580] R.sub.39=C or absent; [1581] R.sub.5,R.sub.67=are independently C, G or absent; [1582] R.sub.2,R.sub.4,R.sub.13,R.sub.15,R.sub.48,R.sub.55=are independently C, G, U or absent; [1583] R.sub.6,R.sub.11,R.sub.22,R.sub.28,R.sub.30,R.sub.34,R.sub.35,R.sub.60,R.sub.61,R.sub.71=are independently C, U or absent; [1584] R.sub.10,R.sub.19,R.sub.51=are independently G or absent; [1585] R.sub.1,R.sub.3,R.sub.20,R.sub.42=are independently G, U or absent; [1586] R.sub.8,R.sub.17,R.sub.21,R.sub.36,R.sub.54=are independently U or absent; [1587] [R.sub.47].sub.x=N or absent;

[1588] wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent.

Variable Region Consensus Sequence

[1589] In an embodiment, a TREM disclosed herein comprises a variable region at position R.sub.47. In an embodiment, the variable region is 1-271 ribonucleotides in length (e.g. 1-250, 1-225, 1-200, 1-175, 1-150, 1-125, 1-100, 1-75, 1-50, 1-40, 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 10-271, 20-271, 30-271, 40-271, 50-271, 60-271, 70-271, 80-271, 100-271, 125-271, 150-271, 175-271, 200-271, 225-271, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, or 271 ribonucleotides). In an embodiment, the variable region comprises any one, all or a combination of Adenine, Cytosine, Guanine or Uracil.

[1590] In an embodiment, the variable region comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 3, e.g., any one of SEQ ID NOs: 452-561 disclosed in Table 3.

TABLE-US-00003 TABLE 3 Exemplary variable region sequences. SEQ ID NO SEQUENCE   1 452 AAAATATAAATATATTTC   2 453 AAGCT   3 454 AAGTT   4 455 AATTCTTCGGAATGT   5 456 AGA   6 457 AGTCC   7 458 CAACC   8 459 CAATC   9 460 CAGC  10 461 CAGGCGGGTTCTGCCCGCGC  11 462 CATACCTGCAAGGGTATC  12 463 CGACCGCAAGGTTGT  13 464 CGACCTTGCGGTCAT  14 465 CGATGCTAATCACATCGT  15 466 CGATGGTGACATCAT  16 467 CGATGGTTTACATCGT  17 468 CGCCGTAAGGTGT  18 469 CGCCTTAGGTGT  19 470 CGCCTTTCGACGCGT  20 471 CGCTTCACGGCGT  21 472 CGGCAGCAATGCTGT  22 473 CGGCTCCGCCTTC  23 474 CGGGTATCACAGGGTC  24 475 CGGTGCGCAAGCGCTGT  25 476 CGTACGGGTGACCGTACC  26 477 CGTCAAAGACTTC  27 478 CGTCGTAAGACTT  28 479 CGTTGAATAAACGT  29 480 CTGTC  30 481 GGCC  31 482 GGGGATT  32 483 GGTC  33 484 GGTTT  34 485 GTAG  35 486 TAACTAGATACTTTCAGAT  36 487 TACTCGTATGGGTGC  37 488 TACTTTGCGGTGT  38 489 TAGGCGAGTAACATCGTGC  39 490 TAGGCGTGAATAGCGCCTC  40 491 TAGGTCGCGAGAGCGGCGC  41 492 TAGGTCGCGTAAGCGGCGC  42 493 TAGGTGGTTATCCACGC  43 494 TAGTC  44 495 TAGTT  45 496 TATACGTGAAAGCGTATC  46 497 TATAGGGTCAAAAACTCTATC  47 498 TATGCAGAAATACCTGCATC  48 499 TCCCCATACGGGGGC  49 500 TCCCGAAGGGGTTC  50 501 TCTACGTATGTGGGC  51 502 TCTCATAGGAGTTC  52 503 TCTCCTCTGGAGGC  53 504 TCTTAGCAATAAGGT  54 505 TCTTGTAGGAGTTC  55 506 TGAACGTAAGTTCGC  56 507 TGAACTGCGAGGTTCC  57 508 TGAC  58 509 TGACCGAAAGGTCGT  59 510 TGACCGCAAGGTCGT  60 511 TGAGCTCTGCTCTC  61 512 TGAGGCCTCACGGCCTAC  62 513 TGAGGGCAACTTCGT  63 514 TGAGGGTCATACCTCC  64 515 TGAGGGTGCAAATCCTCC  65 516 TGCCGAAAGGCGT  66 517 TGCCGTAAGGCGT  67 518 TGCGGTCTCCGCGC  68 519 TGCTAGAGCAT  69 520 TGCTCGTATAGAGCTC  70 521 TGGACAATTGTCTGC  71 522 TGGACAGATGTCCGT  72 523 TGGACAGGTGTCCGC  73 524 TGGACGGTTGTCCGC  74 525 TGGACTTGTGGTC  75 526 TGGAGATTCTCTCCGC  76 527 TGGCATAGGCCTGC  77 528 TGGCTTATGTCTAC  78 529 TGGGAGTTAATCCCGT  79 530 TGGGATCTTCCCGC  80 531 TGGGCAGAAATGTCTC  81 532 TGGGCGTTCGCCCGC  82 533 TGGGCTTCGCCCGC  83 534 TGGGGGATAACCCCGT  84 535 TGGGGGTTTCCCCGT  85 536 TGGT  86 537 TGGTGGCAACACCGT  87 538 TGGTTTATAGCCGT  88 539 TGTACGGTAATACCGTACC  89 540 TGTCCGCAAGGACGT  90 541 TGTCCTAACGGACGT  91 542 TGTCCTATTAACGGACGT  92 543 TGTCCTTCACGGGCGT  93 544 TGTCTTAGGACGT  94 545 TGTGCGTTAACGCGTACC  95 546 TGTGTCGCAAGGCACC  96 547 TGTTCGTAAGGACTT  97 548 TTCACAGAAATGTGTC  98 549 TTCCCTCGTGGAGT  99 550 TTCCCTCTGGGAGC 100 551 TTCCCTTGTGGATC 101 552 TTCCTTCGGGAGC 102 553 TTCTAGCAATAGAGT 103 554 TTCTCCACTGGGGAGC 104 555 TTCTCGAGAGGGAGC 105 556 TTCTCGTATGAGAGC 106 557 TTTAAGGTTTTCCCTTAAC 107 558 TTTCATTGTGGAGT 108 559 TTTCGAAGGAATCC 109 560 TTTCTTCGGAAGC 110 561 TTTGGGGCAACTCAAC

Method of Making TREMs

[1591] Methods for designing and constructing expression vectors and modifying a host cell for production of a target (e.g., a TREM or an enzyme disclosed herein) use techniques known in the art. For example, a cell is genetically modified to express an exogenous TREM using cultured mammalian cells (e.g., cultured human cells), insect cells, yeast, bacteria, or other cells under the control of appropriate promoters. Generally, recombinant methods may be used. See, in general, Pharmaceutical Biotechnology: Fundamentals and Applications, Springer (2013); Green and Sambrook (Eds.), Molecular Cloning: A Laboratory Manual (Fourth Edition), Cold Spring Harbor Laboratory Press (2012). For example, mammalian expression vectors may comprise non-transcribed elements such as an origin of replication, a suitable promoter and enhancer, and other 5′ or 3′ flanking non-transcribed sequences. DNA sequences derived from the SV40 viral genome, for example, SV40 origin, early promoter, enhancer, splice, and polyadenylation sites may be used to provide the other genetic elements required for expression of a heterologous DNA sequence.

[1592] A method of making a TREM or TREM composition disclosed herein comprises use of a host cell, e.g., a modified host cell, expressing a TREM.

[1593] The modified host cell is cultured under conditions that allow for expression of the TREM. In an embodiment, the culture conditions can be modulated to increase expression of the TREM. The method of making a TREM further comprises purifying the expressed TREM from the host cell culture to produce a TREM composition. In an embodiment the TREM is a TREM fragment, e.g., a fragment of a tRNA encoded by a deoxyribonucleic acid sequence disclosed in Table 1. E.g., the TREM includes less than the full sequence of a tRNA, e.g., less than the full sequence of a tRNA with the same anticodon, from the same species as the subject being treated, or both. In an embodiment, the production of a TREM fragment, e.g., from a full length TREM or a longer fragment, can be catalyzed by an enzyme, e.g., an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., RNase A, Dicer, Angiogenin, RNaseP, RNaseZ, Rny1 or PrrC.

[1594] In an embodiment, a method of making a TREM described herein comprises contacting (e.g., transducing or transfecting) a host cell (e.g., as described herein, e.g., a modified host cell) with an exogenous nucleic acid described herein, e.g., a DNA or RNA, encoding a TREM under conditions sufficient to express the TREM. In an embodiment, the exogenous nucleic acid comprises an RNA (or DNA encoding an RNA) that comprises a ribonucleic acid (RNA) sequence of an RNA encoded by a DNA sequence disclosed in Table 1. In an embodiment, the exogenous nucleic acid comprises an RNA sequence (or DNA encoding an RNA sequence) that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identical to an RNA sequence encoded by a DNA sequence provided in Table 1. In an embodiment, the exogenous nucleic acid comprises an RNA sequence (or DNA encoding an RNA sequence) that comprises at least 30 consecutive nucleotides of a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 1. In an embodiment, the exogenous nucleic acid comprises an RNA sequence (or DNA encoding an RNA sequence) that comprises at least 30 consecutive nucleotides of an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identical to an RNA sequence encoded by a DNA sequence provided in Table 1.

[1595] In an embodiment, the host cell is transduced with a virus (e.g., a lentivirus, adenovirus or retrovirus) expressing a TREM, e.g., as described in Example 8.

[1596] The expressed TREM can be purified from the host cell or host cell culture to produce a TREM composition, e.g., as described herein. Purification of the TREM can be performed by affinity purification, e.g., as described in the MACS Isolation of specific tRNA molecules protocol, or other methods known in the art. In an embodiment, a TREM is purified by a method described in Example 7.

[1597] In an embodiment, a method of making a TREM, e.g., a TREM composition, comprises contacting a TREM with a reagent, e.g., a capture reagent comprising a nucleic acid sequence complimentary with a TREM. A single capture reagent or a plurality of capture reagents can be used to make a TREM, e.g., a TREM composition. When a single capture reagent is used, the capture reagent can have at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% complimentary sequence with the TREM. When a plurality of capture reagents is used, a composition of TREMs having a plurality of different TREMs can be made. In an embodiment, the capture reagent can be conjugated to an agent, e.g., biotin.

[1598] In an embodiment, the method comprises denaturing the TREM, e.g., prior to hybridization with the capture reagent. In an embodiment, the method comprises, renaturing the TREM, after hybridization and/or release from the capture reagent.

[1599] In an embodiment, a method of making a TREM, e.g., a TREM composition, comprises contacting a TREM with a reagent, e.g., a separation reagent, e.g., a chromatography reagent. In an embodiment, a chromatography reagent includes a column chromatography reagent, a planar chromatography reagent, a displacement chromatography reagent, a gas chromatography reagent, a liquid chromatography reagent, an affinity chromatography reagent, an ion-exchange chromatography reagent, or a size-exclusion chromatography reagent.

[1600] In an embodiment, a TREM made by any of the methods described herein can be: (i) charged with an amino acid, e.g., a cognate amino acid; (ii) charged with a non-cognate amino acid (e.g., a mischarged TREM (mTREM); or (iii) not charged with an amino acid, e.g., an uncharged TREM (uTREM).

[1601] In an embodiment, a TREM made by any of the methods described herein is an uncharged TREM (uTREM). In an embodiment, a method of making a uTREM comprises culturing the host cell in media that has a limited amount of one or more nutrients, e.g., the media is nutrient starved.

[1602] In an embodiment, a charged TREM, e.g., a TREM charged with a cognate AA or a non-cognate AA, can be uncharged, e.g., by dissociating the AA, e.g., by incubating the TREM at a high temperature.

Exogenous Nucleic Acid Encoding a TREM or a TREM Fragment

[1603] In an embodiment, an exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM comprises a nucleic acid sequence comprising a nucleic acid sequence of one or a plurality of RNA sequences encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, an exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM comprises a nucleic acid sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In one embodiment, the exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM comprises a nucleic acid sequence less than 100% identical to an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1.

[1604] In an embodiment, an exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM comprises the nucleic acid sequence of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, an exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM comprises a nucleic acid sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a plurality of RNA sequences encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, an exogenous nucleic acid encoding a TREM comprises an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, the exogenous nucleic acid encoding a TREM comprises an RNA sequence encoded by a DNA sequence less than 100% identical to a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1.

[1605] In an embodiment, an exogenous nucleic acid, e.g., a DNA or RNA, encoding a TREM comprises an RNA sequence of one or a plurality of TREM fragments, e.g., a fragment of an RNA encoded by a DNA sequence disclosed in Table 1, e.g., as described herein, e.g., a fragment of any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of a nucleic acid sequence of an RNA encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of a nucleic acid sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA encoded by a DNA sequence provided in Table 1. In an embodiment, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of a nucleic acid sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1.

[1606] In an embodiment, a TREM fragment comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 24, 25, 26, 27, 28, 29 or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence disclosed in Table 1 e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 24, 25, 26, 27, 28, 29 or 30 consecutive nucleotides of an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1 e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1. In an embodiment, a TREM fragment comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 24, 25, 26, 27, 28, 29 or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1 e.g., any one of SEQ ID NOs: 1-451 as disclosed in Table 1.

[1607] In an embodiment, the exogenous nucleic acid comprises a DNA, which upon transcription, expresses a TREM.

[1608] In an embodiment, the exogenous nucleic acid comprises an RNA, which upon reverse transcription, results in a DNA which can be transcribed to provide the TREM.

[1609] In an embodiment, the exogenous nucleic acid encoding a TREM comprises: (i) a control region sequence; (ii) a sequence encoding a modified TREM; (iii) a sequence encoding more than one TREM; or (iv) a sequence other than a tRNA.sup.Met sequence.

[1610] In an embodiment, the exogenous nucleic acid encoding a TREM comprises a promoter sequence. In an embodiment, the exogenous nucleic acid comprises an RNA Polymerase III (Pol III) recognition sequence, e.g., a Pol III binding sequence. In an embodiment, the promoter sequence comprises a U6 promoter sequence or fragment thereof. In an embodiment, the nucleic acid sequence comprises a promoter sequence that comprises a mutation, e.g., a promoter-up mutation, e.g., a mutation that increases transcription initiation, e.g., a mutation that increases TFIIIB binding. In an embodiment, the nucleic acid sequence comprises a promoter sequence which increases Pol III binding and results in increased tRNA production, e.g., TREM production.

[1611] Also disclosed herein is a plasmid comprising an exogenous nucleic acid encoding a TREM. In an embodiment, the plasmid comprises a promoter sequence, e.g., as described herein.

TREM Composition

[1612] In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, comprises a pharmaceutically acceptable excipient. Exemplary excipients include those provided in the FDA Inactive Ingredient Database (https://www.accessdata.fda.gov/scripts/cder/iig/index.Cfm).

[1613] In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100 or 150 grams of TREM. In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50 or 100 milligrams of TREM.

[1614] In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% dry weight TREMs.

[1615] In an embodiment, a TREM composition comprises at least 1×10.sup.6 TREM molecules, at least 1×10.sup.7 TREM molecules, at least 1×10.sup.8 TREM molecules or at least 1×10.sup.9 TREM molecules.

[1616] In an embodiment, a TREM composition produced by any of the methods of making disclosed herein can be charged with an amino acid using an in vitro charging reaction as disclosed in Example 11, or as known in the art.

[1617] In an embodiment, a TREM composition comprise one or more species of TREMs. In an embodiment, a TREM composition comprises a single species of TREMs. In an embodiment, a TREM composition comprises a first TREM species and a second TREM species. In an embodiment, the TREM composition comprises X TREM species, wherein X=2, 3, 4, 5, 6, 7, 8, 9, or 10.

[1618] In an embodiment, the TREM has at least 70, 75, 80, 85, 90, or 95, or has 100%, identity with a sequence encoded by a nucleic acid in Table 1.

[1619] In an embodiment, the TREM comprises a consensus sequence provided herein.

[1620] A TREM composition can be formulated as a liquid composition, as a lyophilized composition or as a frozen composition.

[1621] In some embodiments, a TREM composition can be formulated to be suitable for pharmaceutical use, e.g., a pharmaceutical TREM composition. In an embodiment, a pharmaceutical TREM composition is substantially free of materials and/or reagents used to separate and/or purify a TREM, e.g., a separation reagent described herein.

[1622] In some embodiments, a TREM composition can be formulated with water for injection. In some embodiments, a TREM composition formulated with water for injection is suitable for pharmaceutical use, e.g., comprises a pharmaceutical TREM composition.

TREM Purification

[1623] A TREM composition, e.g., a TREM pharmaceutical composition, may be purified from host cells by nucleotide purification techniques. In one embodiment, a TREM composition is purified by affinity purification, e.g., as described in the MACS Isolation of specific tRNA molecules protocol, or by a method described in Example 1-3 or 7. In one embodiment, a TREM composition is purified by liquid chromatography, e.g., reverse-phase ion-pair chromatography (IP-RP), ion-exchange chromatography (IE), affinity chromatography (AC), size-exclusion chromatography (SEC), and combinations thereof. See, e.g., Baronti et al. Analytical and Bioanalytical Chemistry (2018) 410:3239-3252.

[1624] In an embodiment, a TREM composition can be purified with a purification method comprising one, two or all of the following steps, e.g., in the order recited: (i) separating nucleic acids from protein to provide and RNA preparation; (ii) separating RNA with of less than 200 nt from larger RNA species; and/or (iii) separating a TREM from other RNA species by affinity-based separation, e.g., sequence affinity.

[1625] In an embodiment, steps (i)-(iii) are performed in the order recited.

[1626] In an embodiment, the purification method comprises step (i). In an embodiment, step (i) comprises extracting nucleic acids from protein in a sample, e.g., as described in Example 1. In an embodiment, the extraction method comprises a phenol chloroform extraction, In an embodiment, the purification method comprises step (ii). In an embodiment, step (ii) is performed on a sample, after step (i). In an embodiment, step (ii) comprises separating RNA of less than a threshold size, e.g., less than 500 nt, 400 nt, 300 nt, 250 nt, or 200 nt in size from larger RNAs, e.g., using a miRNeasy kit as described in Example 1. In an embodiment, step (ii) comprises performing a salt precipitation, e.g., LiCl precipitation, to enrich for small RNAs (e.g., remove large RNAs), as described in Example 1. In an embodiment, separation of the RNA of less than a threshold size from larger RNAs, e.g., using a miRNeasy kit, is performed prior to the salt precipitation, e.g., LiCl precipitation. In an embodiment, step (ii) further comprises performing a desalting or buffer exchange step, e.g., with a G25 column.

[1627] In an embodiment, the purification method comprises step (iii). In an embodiment, step (iii) comprises performing an affinity-based separation to enrich for a TREM. In an embodiment, step (iii) is performed on a sample after step (i) and/or step (ii). In an embodiment, the affinity based separation comprises a sequence based separation, e.g., using a probe (e.g., oligo) comprising a sequence that binds to a TREM, e.g., as described in Example 1. In an embodiment, the probe (e.g., oligo) comprises one or more tags, e.g., a biotin tag and/or a fluorescent tag.

[1628] In an embodiment, the TREM purification method comprising steps (i), (ii) and (iii) results in a purified TREM composition. In an embodiment, a TREM composition purified according to a method described herein results in lesser RNA contaminants, e.g., as compared to a Trizol RNA extraction purification method.

TREM Quality Control and Production Assessment

[1629] A TREM or a TREM composition, e.g., a pharmaceutical TREM composition, produced by any of the methods disclosed herein can be assessed for a characteristic associated with the TREM or the TREM preparation, such as purity, host cell protein or DNA content, endotoxin level, sterility, TREM concentration, TREM structure, or functional activity of the TREM. Any of the above-mentioned characteristics can be evaluated by providing a value for the characteristic, e.g., by evaluating or testing the TREM, the TREM composition, or an intermediate in the production of the TREM composition. The value can also be compared with a standard or a reference value. Responsive to the evaluation, the TREM composition can be classified, e.g., as ready for release, meets production standard for human trials, complies with ISO standards, complies with cGMP standards, or complies with other pharmaceutical standards. Responsive to the evaluation, the TREM composition can be subjected to further processing, e.g., it can be divided into aliquots, e.g., into single or multi-dosage amounts, disposed in a container, e.g., an end-use vial, packaged, shipped, or put into commerce. In embodiments, in response to the evaluation, one or more of the characteristics can be modulated, processed or re-processed to optimize the TREM composition. For example, the TREM composition can be modulated, processed or re-processed to (i) increase the purity of the TREM composition; (ii) decrease the amount of HCP in the composition; (iii) decrease the amount of DNA in the composition; (iv) decrease the amount of fragments in the composition; (v) decrease the amount of endotoxins in the composition; (vi) increase the in vitro translation activity of the composition; (vii) increase the TREM concentration of the composition; or (viii) inactivate or remove any viral contaminants present in the composition, e.g., by reducing the pH of the composition or by filtration.

[1630] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has a purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, i.e., by mass.

[1631] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has a host cell protein (HCP) contamination of less than 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, 100 ng/ml, 200 ng/ml, 300 ng/ml, 400 ng/ml, or 500 ng/ml.

[1632] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has a host cell protein (HCP) contamination of less than 0.1 ng, 1 ng, 5 ng, 10 ng, 15 ng, 20 ng, 25 ng, 30 ng, 35 ng, 40 ng, 50 ng, 60 ng, 70 ng, 80 ng, 90 ng, 100 ng, 200 ng, 300 ng, 400 ng, or 500 ng per milligram (mg) of the TREM composition.

[1633] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has a DNA content, e.g., host cell DNA content, of less than 1 ng/ml, 5 ng/ml, 10 ng/ml, 15 ng/ml, 20 ng/ml, 25 ng/ml, 30 ng/ml, 35 ng/ml, 40 ng/ml, 50 ng/ml, 60 ng/ml, 70 ng/ml, 80 ng/ml, 90 ng/ml, 100 ng/ml, 200 ng/ml, 300 ng/ml, 400 ng/ml, or 500 ng/ml.

[1634] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has less than 0.1%, 0,5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25% TREM fragments relative to full length TREMs.

[1635] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has low levels or absence of endotoxins, e.g., a negative result as measured by the Limulus amebocyte lysate (LAL) test; In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has in-vitro translation activity, e.g., as measured by an assay described in Example 15.

[1636] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has a TREM concentration of at least 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 0.1 ug/mL, 0.5 ug/mL, 1 ug/mL, 2 ug/mL, 5 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, 40 ug/mL, 50 ug/mL, 60 ug/mL, 70 ug/mL, 80 ug/mL, 100 ug/mL, 200 ug/mL, 300 ug/mL, 500 ug/mL, 1000 ug/mL, 5000 ug/mL, 10,000 ug/mL, or 100,000 ug/mL.

[1637] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) is sterile, e.g., the composition or preparation supports the growth of fewer than 100 viable microorganisms as tested under aseptic conditions, the composition or preparation meets the standard of USP <71>, and/or the composition or preparation meets the standard of USP <85>.

[1638] In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has an undetectable level of viral contaminants, e.g., no viral contaminants. In an embodiment, any viral contaminant, e.g., residual virus, present in the composition is inactivated or removed. In an embodiment, any viral contaminant, e.g., residual virus, is inactivated, e.g., by reducing the pH of the composition. In an embodiment, any viral contaminant, e.g., residual virus, is removed, e.g., by filtration or other methods known in the field.

TREM Administration

[1639] An TREM composition or pharmaceutical composition described herein can be administered to a cell, tissue or subject, e.g., by direct administration to a cell, tissue and/or an organ in vitro, ex-vivo or in vivo. In-vivo administration may be via, e.g., by local, systemic and/or parenteral routes, for example intravenous, subcutaneous, intraperitoneal, intrathecal, intramuscular, ocular, nasal, urogenital, intradermal, dermal, enteral, intravitreal, intracerebral, intrathecal, or epidural.

Vectors and Carriers

[1640] In some embodiments the TREM, or TREM composition described herein, is delivered to cells, e.g. mammalian cells or human cells, using a vector. The vector may be, e.g., a plasmid or a virus. In some embodiments, delivery is in vivo, in vitro, ex vivo, or in situ. In some embodiments, the virus is an adeno associated virus (AAV), a lentivirus, an adenovirus. In some embodiments, the system or components of the system are delivered to cells with a viral-like particle or a virosome. In some embodiments, the delivery uses more than one virus, viral-like particle or virosome.

Carriers

[1641] A TREM, a TREM composition or a pharmaceutical TREM composition described herein may comprise, may be formulated with, or may be delivered in, a carrier.

Viral Vectors

[1642] The carrier may be a viral vector (e.g., a viral vector comprising a sequence encoding a TREM). The viral vector may be administered to a cell or to a subject (e.g., a human subject or animal model) to deliver a TREM, a TREM composition or a pharmaceutical TREM composition. A viral vector may be systemically or locally administered (e.g., injected). Viral genomes provide a rich source of vectors that can be used for the efficient delivery of exogenous genes into a mammalian cell. Viral genomes are known in the art as useful vectors for delivery because the polynucleotides contained within such genomes are typically incorporated into the nuclear genome of a mammalian cell by generalized or specialized transduction. These processes occur as part of the natural viral replication cycle, and do not require added proteins or reagents in order to induce gene integration. Examples of viral vectors include a retrovirus (e.g., Retroviridae family viral vector), adenovirus (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvovirus (e.g., adeno-associated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g., influenza virus), rhabdovirus (e.g., rabies and vesicular stomatitis virus), paramyxovirus (e.g., measles and Sendai), positive strand RNA viruses, such as picornavirus and alphavirus, and double stranded DNA viruses including adenovirus, herpesvirus (e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus, replication deficient herpes virus), and poxvirus (e.g., vaccinia, modified vaccinia Ankara (MVA), fowlpox and canarypox). Other viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, human papilloma virus, human foamy virus, and hepatitis virus, for example. Examples of retroviruses include: avian leukosis-sarcoma, avian C-type viruses, mammalian C-type, B-type viruses, D-type viruses, oncoretroviruses, HTLV-BLV group, lentivirus, alpharetrovirus, gammaretrovirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, Virology (Third Edition) Lippincott-Raven, Philadelphia, 1996). Other examples include murine leukemia viruses, murine sarcoma viruses, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, Gibbon ape leukemia virus, Mason Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentiviruses. Other examples of vectors are described, for example, in U.S. Pat. No. 5,801,030, the teachings of which are incorporated herein by reference. In some embodiments the system or components of the system are delivered to cells with a viral-like particle or a virosome.

Cell and Vesicle-Based Carriers

[1643] A TREM, a TREM composition or a pharmaceutical TREM composition described herein can be administered to a cell in a vesicle or other membrane-based carrier.

[1644] In embodiments, a TREM or TREM composition, or pharmaceutical TREM composition described herein is administered in or via a cell, vesicle or other membrane-based carrier. In one embodiment, the TREM or TREM composition or pharmaceutical TREM composition can be formulated in liposomes or other similar vesicles. Liposomes are spherical vesicle structures composed of a uni- or multilamellar lipid bilayer surrounding internal aqueous compartments and a relatively impermeable outer lipophilic phospholipid bilayer. Liposomes may be anionic, neutral or cationic. Liposomes are biocompatible, nontoxic, can deliver both hydrophilic and lipophilic drug molecules, protect their cargo from degradation by plasma enzymes, and transport their load across biological membranes and the blood brain barrier (BBB) (see, e.g., Spuch and Navarro, Journal of Drug Delivery, vol. 2011, Article ID 469679, 12 pages, 2011. doi:10.1155/2011/469679 for review).

[1645] Vesicles can be made from several different types of lipids; however, phospholipids are most commonly used to generate liposomes as drug carriers. Methods for preparation of multilamellar vesicle lipids are known in the art (see for example U.S. Pat. No. 6,693,086, the teachings of which relating to multilamellar vesicle lipid preparation are incorporated herein by reference). Although vesicle formation can be spontaneous when a lipid film is mixed with an aqueous solution, it can also be expedited by applying force in the form of shaking by using a homogenizer, sonicator, or an extrusion apparatus (see, e.g., Spuch and Navarro, Journal of Drug Delivery, vol. 2011, Article ID 469679, 12 pages, 2011. doi:10.1155/2011/469679 for review). Extruded lipids can be prepared by extruding through filters of decreasing size, as described in Templeton et al., Nature Biotech, 15:647-652, 1997, the teachings of which relating to extruded lipid preparation are incorporated herein by reference.

[1646] Lipid nanoparticles are another example of a carrier that provides a biocompatible and biodegradable delivery system for the TREM or TREM compositions or pharmaceutical TREM composition described herein. Nanostructured lipid carriers (NLCs) are modified solid lipid nanoparticles (SLNs) that retain the characteristics of the SLN, improve drug stability and loading capacity, and prevent drug leakage. Polymer nanoparticles (PNPs) are an important component of drug delivery. These nanoparticles can effectively direct drug delivery to specific targets and improve drug stability and controlled drug release. Lipid-polymer nanoparticles (PLNs), a new type of carrier that combines liposomes and polymers, may also be employed. These nanoparticles possess the complementary advantages of PNPs and liposomes. A PLN is composed of a core-shell structure; the polymer core provides a stable structure, and the phospholipid shell offers good biocompatibility. As such, the two components increase the drug encapsulation efficiency rate, facilitate surface modification, and prevent leakage of water-soluble drugs. For a review, see, e.g., Li et al. 2017, Nanomaterials 7, 122; doi:10.3390/nano7060122.

[1647] Exosomes can also be used as drug delivery vehicles for the TREM or TREM compositions or pharmaceutical TREM composition described herein. For a review, see Ha et al. July 2016. Acta Pharmaceutica Sinica B. Volume 6, Issue 4, Pages 287-296; https://doi.org/10.1016/j.apsb.2016.02.001.

[1648] Ex vivo differentiated red blood cells can also be used as a carrier for a TREM or TREM composition, or pharmaceutical TREM composition described herein. See, e.g., WO2015073587; WO2017123646; WO2017123644; WO2018102740; wO2016183482; WO2015153102; WO2018151829; WO2018009838; Shi et al. 2014. Proc Natl Acad Sci USA. 111(28): 10131-10136; U.S. Pat. No. 9,644,180; Huang et al. 2017. Nature Communications 8: 423; Shi et al. 2014. Proc Natl Acad Sci USA. 111(28): 10131-10136.

[1649] Fusosome compositions, e.g., as described in WO2018208728, can also be used as carriers to deliver the TREM or TREM composition, or pharmaceutical TREM composition described herein.

Use of TREMs

[1650] A TREM composition (e.g., a pharmaceutical TREM composition described herein) can modulate a function in a cell, tissue or subject. In embodiments, a TREM composition (e.g., a pharmaceutical TREM composition) described herein is contacted with a cell or tissue, or administered to a subject in need thereof, in an amount and for a time sufficient to modulate (increase or decrease) one or more of the following parameters: adaptor function (e.g., cognate or non-cognate adaptor function), e.g., the rate, efficiency, robustness, and/or specificity of initiation or elongation of a polypeptide chain; ribosome binding and/or occupancy; regulatory function (e.g., gene silencing or signaling); cell fate; mRNA stability; protein stability; protein transduction; protein compartmentalization. A parameter may be modulated, e.g., by at least 5% (e.g., at least 10%, 15%, 20%, 25%, 30%, 40%. 50%. 60%. 70%, 80%, 90%, 100%, 150%, 200% or more) compared to a reference tissue, cell or subject (e.g., a healthy, wild-type or control cell, tissue or subject).

[1651] All references and publications cited herein are hereby incorporated by reference.

[1652] The following examples are provided to further illustrate some embodiments of the present invention, but are not intended to limit the scope of the invention; it will be understood by their exemplary nature that other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used.

EXAMPLES

[1653]

TABLE-US-00004 Table of Contents for Examples Manufacture and preparation of TREMs Example 1 Manufacture of a TREM in a mammalian production host cell from transient transfection Example 2 Manufacture of a TREM in a mammalian production host cell from stable cell lines Example 3 Manufacture of a TREM in a mammalian production host cell from stable cell lines Delivery of TREMs Example 4 Delivery of TREMs to mammalian cells Assays to analyze TREM activity Example 5 TREM functional activity assay in mammalian cells Example 6 TREM translational activity assay in Human Cell Extract Cell-Free Protein Synthesis (hCFPS) lysate Manufacture and preparation of TREMs Example 7 Manufacture of a TREM in a mammalian production host cell, and use thereof to modulate a cellular function −1 Example 8 Manufacture of a TREM in a mammalian production host cell, and use thereof to modulate a cellular function −2 Example 9 Manufacture of a TREM in modified mammalian production host cell expressing an oncogene Example 10 Preparation of a TREM production host cell modified to inhibit a repressor of tRNA synthesis Example 11 Manufacture of a TREM in modified mammalian production host cell overexpressing an oncogene and a tRNA modifying enzyme Production of TREMs Example 12 Production of a mischarged TREM Example 13 Production of a TREM fragment (in vitro) Example 14 Production of a TREM fragment in a cell expression system Assays to analyze TREM activity Example 15 TREM translational activity assay Example 16 Assay for modulation of cell state Example 17 Assay for the activity of an uncharged TREM to modulate autophagy Example 18 Assay for activity of a mischarged TREM (mTREM)

Example 1: Manufacture of a TREM in a Mammalian Production Host Cell from Transient Transfection

[1654] This example describes the manufacture of a TREM produced in mammalian host cells which transiently express a TREM.

Plasmid Generation

[1655] To generate a plasmid comprising a sequence encoding a TREM, in this example, iMet-CAT TREM, a DNA fragment containing one copy of the sequence AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTCGATGGATCG AAACCATCCTCTGCTA (SEQ ID NO: 262) was synthesized and cloned into the pLKO.1-puro-mCherry backbone plasmid with a U6 promoter following the manufacturer's instructions and standard molecular cloning techniques.

Transfection

[1656] Three (3) μg of plasmid described above was used to transfect a T175 flask of HEK293T cells plated at 80% confluency using 9 uL of lipofectamine RNAiMax reagents according to the manufacturer's instructions. Cells were harvested at 48 hours post-transfection for purification.

Purification Using a Small RNA Isolation Kit

[1657] The iMet-overexpressing cells were lysed. To generate a small RNA (sRNA) fraction, a small RNA isolation kit, such as the Qiagen miRNeasy kit, was used to separate RNAs smaller than 200 nucleotides from the rest of the total RNA pool in the lysate, per manufacturer's instructions. To further exclude larger RNAs, a LiCl precipitation was performed to remove remaining large RNAs in the sRNA fraction. Finally, the sRNA fraction was added to a G50 column to remove RNAs smaller than 10 nucleotides from the sRNA fraction and for buffer exchange.

[1658] To isolate the TREM from the sRNA fraction, a probe binding method was used. A biotinylated capture probe corresponding to a DNA probe or a 2′-OMe nucleic acid that is complementary to a unique region of the target TREM being purified, in this example, a probe conjugated to biotin at the 5′ end with the sequence TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 267), was used to bind and purify the iMet-CAT-TREM. The sRNA fraction was incubated with annealing buffer and the biotinylated capture probe at 90° C. for 4-5 minutes and cooled at a rate of 0.1° C./s to 25° C.

[1659] The admixture was then incubated with binding buffer and streptavidin-conjugated RNase-free magnetic beads for 15 minutes to enable binding of the DNA-TREM complexes to the beads. The mixture was then added to a magnetic field separator rack and washed 2-3 times with wash buffer. The TREM retained on the beads was eluted by adding elution buffer with or without a DNase enzyme to ensure complete removal of the DNA capture probe and then admixed with a pharmaceutically acceptable excipient to make a test TREM product.

Example 2: Manufacture of a TREM in a Mammalian Production Host Cell from Stable Cell Lines

[1660] This example describes the manufacture of a TREM produced in mammalian host cells stably expressing a TREM.

Preparation of TREM Expressing Lentivirus

[1661] To prepare a TREM expressing lentivirus in a 10 mm dish, packaging cells, such as HEK293T cells (293T cells (ATCC® CRL-3216™), were forward transfected with 9 μg of a plasmid comprising a sequence encoding a TREM as described in Example 1, and 9 μg ViraPower lentiviral packaging mix using TransIT-LT1 transfection reagents according to the manufacturer's instructions.

[1662] After 18 hours, the media was replaced with fresh antibiotic-free high-FBS (30% FBS) media and 24 hours later, the media containing the virus was harvested and stored at 4° C. Another 15 mL of high-FBS media was added to the plate and harvested 24 hours later. Both virus-containing media harvests were pooled and filtered through a 0.45-micron filter. The viral copy number was assessed using the Lenti-X qRT-PCR Titration Kit according to the manufacturer's protocol.

Transduction of Host Cells with TREM Expressing Lentivirus

[1663] To transduce the cells with TREM expressing lentivirus, the lentivirus-containing media was diluted with complete cell media at a 1:4 ratio, in the presence of 10 μg/mL polybrene, and added to the cells. In this example 293T cells were used. The plate was spun for 2 hours at 1000×g to spin infect the cells. After 18 hours, the media was replaced to allow the cells to recover. Forty-eight hours after transduction, puromycin (at 2 μg/mL) antibiotic selection was performed for 5-7 days alongside a population of untransduced control cells.

[1664] The TREMs were isolated, purified, and formulated as described in Example 1 to result in a TREM preparation.

Purification Using Phenol Chloroform Extraction

[1665] The total RNA pool from cells was recovered from cells by guanidinium thiocyanate-phenol-chloroform extraction and concentrated by ethanol precipitation as described in J. Sambrook and D. Russell (2001) Molecular Cloning: A Laboratory Manual, vol. 2, Cold Spring Harbor Laboratory Press, New York, N.Y., USA, 3rd edition 2. The total tRNA pool in the precipitate was then separated from larger nucleic acids (including rRNA and DNA) by precipitation under high lithium salt conditions as described in Cathala, G. et al., DNA, 1983; 2(4):329-35. The elution fraction containing the TREM was further purified through probe binding.

[1666] The TREM fraction was incubated with annealing buffer and the biotinylated capture probe corresponding to a DNA probe or a 2′-OMe nucleic acid that is complementary to a unique region of the target TREM being purified. In this example, a probe conjugated to biotin at the 5′ end with the sequence TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 267), was used to purify the TREM comprising iMet-CAT. The mixture was incubated at 90° C. for 4-5 minutes and cooled at a rate of 0.1° C./s to 25° C.

[1667] The admixture was then incubated with binding buffer and streptavidin-conjugated RNase-free magnetic beads for 15 minutes to enable binding of the DNA-TREM complexes to the beads. The mixture was then added to a magnetic field separator rack and washed 2-3 times. The TREM retained on the beads were eluted by adding elution buffer with or without a DNase enzyme to ensure complete removal the DNA capture probe and then admixed with a pharmaceutically acceptable excipient to make a test TREM product.

Example 3: Manufacture of a TREM in a Mammalian Production Host Cell from Stable Cell Lines

[1668] This example describes the manufacture of a TREM from crude cell lysate, produced from mammalian host cells.

Generation of Stable Cells Expressing TREM

[1669] In this example, a plasmid comprising a sequence encoding a TREM is generated as described in Example 1 or 2. Preparation of TREM expressing lentivirus and transduction of host cells with TREM-expressing lentivirus was performed as described in Example 2.

Purification from Crude Cell Lysate

[1670] The TREM-overexpressing cells, in this example the iMet-CAT-TREM overexpressing cells, were lysed and the lysed material was incubated with annealing buffer and the biotinylated capture probe corresponding to a DNA probe or a 2′-OMe nucleic acid that is complementary to a unique region of the target TREM being purified. In this example, a probe conjugated to biotin at the 5′ end with the sequence TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 267), was used to purify the TREM comprising iMet-CAT. The mixture was incubated at 90° C. for 4-5 minutes and cooled at a rate of 0.1° C./s to 25° C.

[1671] The admixture was then incubated with binding buffer and streptavidin-conjugated RNase-free magnetic beads for 15 minutes to enable binding of the DNA-TREM complexes to the beads. The mixture was then added to a magnetic field separator rack and washed 2-3 times. The TREM retained on the beads were eluted by adding elution buffer with or without a DNase enzyme to ensure complete removal the DNA capture probe and then admixed with a pharmaceutically acceptable excipient to make a test TREM product.

Example 4: Delivery of TREMs to Mammalian Cells

[1672] This example describes the delivery of a TREM to mammalian cells.

[1673] To ensure proper folding, the TREM was heated at 85° C. for 2 minutes and then snap cooled at 4° C. for 5 minutes. To deliver the TREM to mammalian cells, 100 nM of two TREM preparations labeled with Cy3 at different positions (Cy3-iMET-1 and Cy3-iMET-2) were transfected in U2OS (U-2 OS (ATCC® HTB-96™)), H1299 (NCI-H1299 (ATCC® CRL-5803™)), and HeLa (HeLa (ATCC® CCL-2™)) cells using RNAiMax reagents according to the manufacturer's instructions. After 18 hours, the transfection media was removed and replaced with fresh complete media (U2OS: McCoy's 5A, 10% FBS, 1% PenStrep; H1299: RPMI1640, 10% FBS, 1% PenStrep; HeLa: EMEM, 10% FBS, 1% PenStrep).

[1674] To observe TREM delivery to cells, the cells were monitored in a live cell analysis system. In this example, the IncuCyte (from Essen Bioscience) was used to monitor cells. The cells were monitored for 4 days (20×, red 550 ms).

[1675] Cy3 fluorescence signal was readily detected from cells that had been delivered the Cy3-labeled TREMs. The Cy3 fluorescence signal was observed for over 48 hours from the cells in which the TREMs had been delivered. Detection of Cy-3 fluorescence from the cells confirmed delivery of the Cy3-labeled TREM to the cells.

Example 5: Increased Cell Growth in Mammalian Cells with TREM

[1676] This example describes increased cell growth of a mammalian cell upon TREM delivery.

[1677] To ensure proper folding, the iMet TREM was heated at 85° C. for 2 minutes and then snap cooled at 4° C. for 5 minutes. To deliver the iMet TREM to mammalian cells, 100 nM of Cy3-labeled iMet TREM was transfected in U2OS (U-2 OS (ATCC® HTB-96™)), H1299 (NCI-H1299 (ATCC® CRL-5803™)), and HeLa (HeLa (ATCC® CCL-2™)) cells using RNAiMax reagents according to the manufacturer's instructions. As a control, a Cy3-labeled non targeted control siRNA was delivered to cells. After 18 hours, the transfection media was removed and replaced with fresh complete media (U2OS: McCoy's 5A, 10% FBS, 1% PenStrep; H1299: RPMI1640, 10% FBS, 1% PenStrep; HeLa: EMEM, 10% FBS, 1% PenStrep). To observe changes in cell growth, the cells were monitored in a live cell analysis system, in this example in the IncuCyte (from Essen Bioscience), for 4 days (20×, phase contrast).

[1678] Delivery of iMet TREM to U2OS cells (FIG. 1A), H1299 (FIG. 1B) or Hela cells (FIG. 1C) led to a substantial increase in cell growth in all of the cell lines that were tested. The increase in cell growth was compared to cell growth observed with delivery of a Cy3-labeled non-targeted control (Cy3-NTC). The data demonstrates that delivery of a TREM to cells results in increased proliferation and growth.

Example 6: TREM Translational Activity Assay in Human Cell Extract Cell-Free Protein Synthesis (hCFPS) Lysate

[1679] This example describes a TREM mediated increase in translational activity in a cell-free lysate system.

Preparing Human Cell Extracts

[1680] HEK293T cells were grown to ˜80% confluency in 40×150 mm culture dishes. The cells were harvested, washed in PBS, resuspended 1:1 in ice-cold hypotonic lysis buffer (20 mM HEPES pH 7.6, 10 mM KAc, 1.5 mM MgAc, 5 mM DTT and 5× complete EDTA-free proteinase inhibitor cocktail) and incubated on ice for 30 minutes. Cells were lysed using a Dounce homogenizer or by passing the lysate through a 27 G needle, until >95% of the cells were disrupted. The lysate was centrifuged at 14,000 g for 10 mins at 4° C., the supernatant was collected and diluted with the hypotonic lysis buffer to get a ˜15 mg/ml protein solution.

Transcribing mRNAs

[1681] mRNA transcription templates were designed to have a T7 polymerase promoter, a beta-globin 3′UTR, a nanoLuc ORF, and a short artificial 3′UTR. The templates were PCR amplified and used to transcribe capped and poly-adenylated mRNAs with a HiScribe T7 ARCA mRNA kit with tailing (New England Biolabs) following the manufacturer's recommended protocol.

[1682] Performing the TREM Translational Activity Assay in hCFPS Lysate

[1683] Translation reactions were set up in translation buffer (16 mM HEPES pH 7.6, 2.2 mM MgAc, 60 mM KCl, 0.02 mM complete amino acid mix, 1 mM ATP, 0.5 mM GTP, 20 mM creatine phosphate, 0.1 μg/L creatine kinase, 0.1 mM spermidine, 2 U/μl RiboLock RNase Inhibitor) with 35% HEK293T lysate, 0.02 μM capped and poly-adenylated nanoLuc mRNA and 2 μM cell-purified TREM (purified according to Example 2). The reactions were performed in 10 μl triplicates at 37° C. for 30 minutes. For the control reactions, one control reaction was performed with no TREM addition to the reaction and one control reaction was performed with no mRNA addition to the reaction. Then, the NanoLuc activity was detected by mixing each reaction with 40 μl of room temperature Nano-Glo Luciferase assay system (Promega) and reading the luminescence in a plate reader.

[1684] As shown in FIG. 2, the iMET TREM reaction resulted in about a 1.5 fold increase in NanoLuc expression as compared to the control reaction (buffer). The data shows that delivery of the TREM results in an increase in nanoLuc mRNA translation as reflected by an increase in luminescence.

Example 7: Manufacture of a TREM in a Mammalian Production Host Cell, and Use Thereof to Modulate a Cellular Function-1

[1685] This example describes the manufacture of a TREM produced in mammalian host cells.

Plasmid Generation

[1686] To generate a plasmid comprising a sequence encoding a TREM, in this example, iMet-CAT TREM, a DNA fragment with genomic location 6p22.2 and sequence AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTCGATGGATCG AAACCATCCTCTGCTA (SEQ ID NO: 622)) is PCR-amplified from human genomic DNA using the following primer pairs: 5′-TGAGTTGGCAACCTGTGGTA (SEQ ID NO: 623) and 5′-TTGGGTGTCCATGAAAATCA (SEQ ID NO: 624). This fragment is cloned into the pLKO.1 puro backbone plasmid with a U6 promoter (or any other RNA polymerase III recruiting promoter) following the manufacturer's instructions.

Transfection

[1687] One (1) mg of plasmid described above is used to transfect a 1 L culture of suspension-adapted HEK293T cells (Freestyle 293-F cells) at 1×10.sup.5 cells/mL. Cells are harvested at 24, 48, 72, or 96 hours post-transfection to determine the optimized timepoint for TREM expression as determined by Northern blot, or by quantitative PCR (q-PCR).

Purification

[1688] At the optimized harvest cell density point, the TREM is purified as previously described in Cayama et al., Nucleic Acids Research. 28 (12), e64 (2000). Briefly, short RNAs (e.g., tRNAs) are recovered from cells by phenol extraction and concentrated by ethanol precipitation. The total tRNA in the precipitate is then separated from larger nucleic acids (including rRNA and DNA) under high salt conditions by a stepwise isopropanol precipitation. The elution fraction containing the TREM is further purified through probe binding. The TREM fraction is incubated with annealing buffer and the biotinylated capture probe corresponding to a DNA probe or a 2′-OMe nucleic acid that is complementary to a unique region of the target TREM being purified, in this example, a probe conjugated to biotin at the 3′ end with the sequence UAGCAGAGGAUGGUUUCGAUCCAUCA (SEQ ID NO: 625), is used to purify the iMet-CAT-TREM. The mixture is incubated at 90° C. for 2-3 minutes and quickly cooled down to 45° C. and incubated overnight at 45° C. The admixture is then incubated with binding buffer previously heated to 45° C. and streptavidin-conjugated RNase-free magnetic beads for 3 hours to allow binding of the DNA-TREM complexes to the beads. The mixture is then added to a pre-equilibrated column in a magnetic field separator rack and washed 4 times. The TREM retained on the beads are eluted three times by adding elution buffer pre-heated to 80° C. and then admixed with a pharmaceutically acceptable excipient to make a test TREM product.

Use

[1689] One microgram of the test TREM preparation and a control agent are contacted by transfection, electroporation or liposomal delivery, with a cultured cell line, such as a HEP-3B or HEK293T, a tissue or a subject, for a time sufficient for the TREM preparation to modulate a translation level or activity of the cell, relative to the control agent.

Example 8: Manufacture of a TREM in a Mammalian Production Host Cell, and Use Thereof to Modulate a Cellular Function-2

[1690] This example describes the manufacture of a TREM produced in mammalian host cells.

Plasmid Generation

[1691] To generate a plasmid comprising a sequence encoding a TREM, in this example, iMet-CAT-TREM, a DNA fragment containing at least one copy of the sequence AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTCGATGGATCG AAACCATCCTCTGCTA (SEQ ID NO: 626) is synthesized and cloned into the pLKO.1 puro backbone plasmid with a U6 promoter (or any other RNA polymerase III recruiting promoter) following the manufacturer's instructions and standard molecular cloning techniques.

Transfection

[1692] One (1) mg of plasmid described above is used to transfect a 1 L culture of suspension-adapted HEK293T cells (Freestyle 293-F cells) at 1×10.sup.5 cells/mL. Cells are harvested at 24, 48, 72, or 96 hours post-transfection to determine the optimized timepoint for TREM expression as determined by Northern blot, or by quantitative PCR (q-PCR) or Nanopore sequencing.

Purification

[1693] At the optimized harvest timepoint, the cells are lysed and separation from the lysate of RNAs smaller than 200 nucleotides is performed using a small RNA isolation kit per manufacturer's instructions, to generate a small RNA (sRNA) fraction.

[1694] To prepare the affinity purification reagents, streptavidin-conjugated RNase-free magnetic beads are incubated at room temperature for 30 min with 200 mM of biotinylated oligonucleotides corresponding to a DNA probe or a 2′-OMe nucleic acid that is complementary to a unique region of the target TREM being purified. In this example, a probe with the sequence 5′biotin-TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 627) is used to purify the -iMet-CAT-TREM. The beads are washed and heated for 10 min at 75° C.

[1695] The sRNA fraction is heated for 10 min at 75° C. and then mixed with the affinity purification reagent described above. The admixture is incubated at room temperature for 3 hours to allow binding of the TREMs to the bead-bound DNA probe in a sequence specific manner. The beads are then washed until the absorbance of the wash solution at 260 nm is close to zero. Alternatively, the beads are washed three times and the final wash is examined by UV spectroscopy to measure the amount of nucleic acid present in the final wash. The TREM retained on the beads are eluted three times using RNase-free water which can be pre-heated to 80° C., and then admixed with a pharmaceutically acceptable excipient to make a test TREM product.

Use

[1696] One microgram of the test TREM preparation and a control agent are contacted by transfection, electroporation or liposomal delivery, with a cultured cell line, such as HeLa, HEP-3B or HEK293T, a tissue or a subject, for a time sufficient for the TREM preparation to modulate a translation level or activity of the cell, relative to the control agent.

Example 9: Manufacture of a TREM in Modified Mammalian Production Host Cell Expressing an Oncogene

[1697] This example describes the manufacture of a TREM in mammalian host cells modified to overexpress Myc.

Plasmid Generation and Host Cell Modification

[1698] To make the production host cells for this example, HeLa cells (ATCC® CCL-2™) or HEP-3B cells (ATCC® HB-8064™) are transfected with a plasmid containing the gene sequence coding for the c-myc oncogene protein (e.g., pcDNA3-cmyc (Addgene plasmid #16011)) using routine molecular biology techniques. The resulting cell line is referred to herein as HeLamyc+ host cells or HEP-3Bmyc+ host cells.

Preparation of TREM Expressing Lentivirus

[1699] To prepare a TREM expressing lentivirus, HEK293T cells are co-transfected with 3 μg of each packaging vector (pRSV-Rev, pCMV-VSVG-G and pCgpV) and 9 μg of the plasmid comprising a sequence encoding a TREM as described in Example 7, using Lipofectamine 2000 according to manufacturer's instructions. After 24 hours, the media is replaced with fresh antibiotic-free media and after 48 hours, virus-containing supernatant is collected and centrifuged for 10 min at 2000 rpm before being filtered through a 0.45 m filter.

Transduction of Host Cells with TREM Expressing Lentivirus

[1700] Two (2) mL of virus prepared as described above is used to transduce 100,000 HeLamyc+ host cells or HEP-3Bmyc+ host cells, in the presence of 8 μg/mL polybrene. Forty-eight hours after transduction, puromycin (at 2 μg/mL) antibiotic selection is performed for 2-7 days alongside a population of untransduced control cells.

[1701] The TREMs are isolated, purified, and formulated as described in Example 7 or 8 to result in a TREM composition or preparation.

Example 10: Preparation of a TREM Production Host Cell Modified to Inhibit a Repressor of tRNA Synthesis

[1702] This example describes the preparation of Hek293Maf-/TRM1 cells for the production of a TREM.

[1703] Maf1 is a repressor of tRNA synthesis. A Maf1 knockout HEK293T cell line is generated using standard CRISPR/Cas knockout techniques, e.g., a CRISPR/Cas system can be designed to introduce a frameshift mutation in a coding exon of Maf1 to reduce the expression of Maf1 or knockout Maf1 expression, to generate a Hek293Maf-cell line that has reduced expression level and/or activity of Maf1. This cell line is then transfected with an expression plasmid for modifying enzyme Trm1 (tRNA (guanine26-N2)-dimethyltransferase) such as pCMV6-XL4-Trm1, and selected with a selection marker, e.g., neomycin, to generate a stable cell line overexpressing Trm1 (Hek293Maf-/TRM1 cells).

[1704] Hek293Maf-/TRM1 cells can be used as production host cells for the preparation of a TREM as described in any of Examples 7-9.

Example 11: Manufacture of a TREM in Modified Mammalian Production Host Cells Overexpressing an Oncogene and a tRNA Modifying Enzyme

[1705] This Example describes the manufacture of a TREM in mammalian host cells modified to overexpress Myc and Trm1.

Plasmid Generation

[1706] In this example, a plasmid comprising a TREM is generated as described in Example 7 or 8.

Host Cell Modification, Transduction and Purification

[1707] A human cell line, such as HEK293T, stably overexpressing Myc oncogene is generated by transduction of retrovirus expressing the myc oncogene from the pBABEpuro-c-myc.sup.T58A plasmid into HEK293T cells. To generate myc-expressing retrovirus, HEK293T cells are transfected using the calcium phosphate method with the human c-myc retroviral vector, pBABEpuro-c-myc.sup.T58A and the packaging vector, ψ2 vector. After 6 hours, transfection media is removed and replaced with fresh media. After a 24-hour incubation, media is collected and filtered through a 0.45 um filter. For the retroviral infection, HEK293T cells are infected with retrovirus and polybrene (8 ug/ml) using spin infection at 18° C. for 1 hour at 2500 rpm. After 24 hours, the cell culture medium is replaced with fresh medium and 24 hours later, the cells are selected with 2 μg/mL puromycin. Once cells stably overexpressing the oncogene myc are established, they are transfected with a Trm1 plasmid, such as the pCMV6-XL4-Trm1 plasmid, and selected with a selection marker, in this case with neomycin, to generate a stable cell line overexpressing Trm1, in addition to Myc. In parallel, lentivirus to overexpress TREM is generated as described in Example 9 with HEK293T cells and PLKO.1-TREM vectors.

[1708] One hundred thousand (1×10.sup.5) cells overexpressing Myc and Trm1 are transduced with the TREM virus in the presence of 8 μg/mL polybrene. Media is replaced 24 hours later. Forty-eight hours after transduction, antibiotic selection is performed with 2 μg/mL puromycin for 2-7 days alongside a population of untransduced control cells. The TREMs are isolated, purified and formulated using the method described in Example 7 or 8 to produce a TREM preparation.

Example 12: Production of a Mischarged TREM

[1709] This example describes the production of a TREM charged with an amino acid that does not correspond to its natural anticodon.

[1710] A TREM is produced as described in any of Examples 7-11. The TREM product is charged with a heterologous amino acid using an in vitro charging reaction known in the art (see, e.g., Walker & Fredrick (2008) Methods (San Diego, Calif.) 44(2):81-6). Briefly, the purified TREM, for example a TREM comprising tRNA-Val(GTG), is placed in a buffer with the heterologous amino acid of interest (for example glutamic acid), and the corresponding aminoacyl-tRNA synthetase (for example a Valyl-tRNA synthetase mutated to enhance tRNA mischarging), to induce TREM charging.

[1711] To isolate the aminoacyl-TREM, the in vitro charging reaction is passed through a spin column and the concentration based on the A.sub.260 absorbance is determined as is the extent of aminoacylation using acid gel electrophoresis. Aminoacylated TREM can also be isolated by binding to His6-tagged EF-Tu (“His6” disclosed as SEQ ID NO: 628), followed by affinity chromatography on Ni-NTA agarose, phenol-chloroform extraction and subsequent precipitation of the nucleic acids as described in Rezgui et al., 2013, PNAS 110:12289-12294.

Example 13: Production of a TREM Fragment (In Vitro)

[1712] This example describes the production of a TREM fragment in vitro, from a TREM manufactured in mammalian host cells.

[1713] A TREM is made as described in any one of Examples 7-13 above. An enzymatic cleavage assay with enzymes known to generate tRNA fragments, such as RNase A or angiogenin, is used to produce fragments for administration to a cell, tissue or subject.

[1714] Briefly, a TREM manufactured as describe above is incubated in one of: 0.1M Hepes/NaOH, pH 7.4 with 10 nM final concentration of RNase A for 10 min at 30° C., or 0.1M MES, 0.1M NaCl, pH 6.0, with an effective amount of angiogenin, and BSA for 6 hours at 37° C.

[1715] To isolate a target TREM fragment after enzymatic treatment, a sequence affinity purification procedure is performed, as described above.

Example 14: Production of a TREM Fragment in a Cell Expression System

[1716] This example describes the production of a TREM fragment in a cell expression system.

[1717] A cell line stably overexpressing a TREM is generated as described in any of Examples 7-9 or 11. Hek293T cells overexpressing the TREM are treated with 0.5 pg/ml recombinant angiogenin for 90 min before total RNA is extracted with Trizol. Size selection of RNAs smaller than 200 nucleotides is performed using a small RNA isolation kit per manufacturer's instructions. Streptavidin-conjugated RNase-free magnetic beads are incubated at room temperature for 30 min with 200 mM of biotinylated oligonucleotides corresponding to a probe or a DNA probe that is complementary to a unique region of the TREM fragment being purified. The beads are washed and heated for 10 min at 75° C. The size-selected RNA eluate is also heated for 10 min at 75° C. and then mixed with the beads. The TREM-bead mixture is incubated at room temperature for 3 hours to allow binding of the TREMs to the bead-bound DNA probe. The beads are then washed until the wash solution at 260 nm is close to zero (0). Alternatively, the beads are washed three times and the final wash is examined by UV spectroscopy to measure the amount of nucleic acid present in the final wash. The TREM retained on the beads are eluted 3 times using RNase-free water pre-heated to 80° C. or elution buffer pre-heated to 80° C.

Example 15: TREM Translational Activity Assays

[1718] This example describes assays to evaluate the ability of a TREM to be incorporated into a nascent polypeptide chain.

Translation of the FLAG-AA-his Peptide Sequence

[1719] A test TREM is assayed in an in-vitro translation reaction with an mRNA encoding the peptide FLAG-XXX-His6× (“His6” disclosed as SEQ ID NO: 628), where XXX are 3 consecutive codons corresponding to the test TREM anticodon.

[1720] A tRNA-depleted rabbit reticulocyte lysate (Jackson et al. 2001. RNA 7:765-773) is incubated 1 hour at 30° C. with 10-25 ug/mL of the test TREM in addition to 10-25 ug/mL of the tRNAs required for the FLAG and His tag translation. In this example, the TREM used is Ile-GAT-TREM, therefore the peptide used is FLAG-LLL-His6× (“His6” disclosed as SEQ ID NO: 628) and the TREM added is TREM-Ile-GAT, in addition to the following, which are added to translate the peptide FLAG and HIS tags: tRNA-Asp-GAC, tRNA-Tyr-TAC, tRNA-Lys-AAA, tRNA-Lys-AAAG, tRNA-Asp-GAT, tRNA-His-CAT. To determine if the test TREM is functionally able to be incorporated into a nascent peptide, an ELISA capture assay is performed. Briefly, an immobilized anti-His6× antibody (“His6” disclosed as SEQ ID NO: 628) is used to capture the FLAG-LLL-His6× peptide (“His6” disclosed as SEQ ID NO: 628) from the reaction mixture. The reaction mixture is then washed off and the peptide is detected with an enzyme-conjugated anti-FLAG antibody, which reacts to a substrate in the ELISA detection step. If the TREM produced is functional, the FLAG-LLL-His6 peptide (“His6” disclosed as SEQ ID NO: 628) is produced and detection occurs by the ELISA capture assay.

[1721] If the TREM produced is not functional, the FLAG-LLL-His6 peptide (“His6” disclosed as SEQ ID NO: 628) is not produced and no detection occurs by the ELISA capture assay.

Translational Suppression Assay

[1722] This assay describes a test TREM having translational adaptor molecule function by rescuing a suppression mutation and allowing the full protein to be translated. The test TREM, in this example Ile-CUA-TREM, is produced such that it contains the sequence of the Ile-GAT-TREM body but with the anticodon sequence corresponding to CUA instead of GAT. HeLa cells are co-transfected with 50 ng of TREM and with 200 ng of a DNA plasmid encoding a mutant GFP containing a TAG stop codon at the S29 position as described in Geslain et al. 2010. J Mol Biol. 396:821-831. HeLa cells transfected with the GFP plasmid alone serve as a negative control. After 24 hours, cells are collected and analyzed for fluorescence recovery by flow cytometry. The fluorescence is read out with an emission peak at 509 nm (excitation at 395 nm). It is expected that if the test TREM is functional, it can or will be sufficient to rescue the stop mutation in the GFP molecule and can produce the full-length fluorescent protein, which is detected by flow cytometry. If the test TREM is not functional or is less functional, the stop mutation is likely not to be rescued, and no fluorescence is emitted from the GFP molecule and accordingly a reduced GFP signal or no GFP signal is detected by flow cytometry.

In Vitro Translational Assay

[1723] This assay describes a test TREM having translational adaptor molecule function by successfully being incorporated into a nascent polypeptide chain in an in vitro translation reaction. First, a rabbit reticulocyte lysate that is depleted of the endogenous tRNA using an antisense or complimentary oligonucleotide which (i) targets the sequence between the anticodon and variable loop; or (ii) binds the region between the anticodon and variable loop is generated (see, e.g., Cui et al. 2018. Nucleic Acids Res. 46(12):6387-6400). 10-25 ug/mL of the test TREM is added in addition to 2 ug/uL of a GFP-encoding mRNA to the depleted lysate. A non-depleted lysate with the GFP mRNA, with or without the test TREM added are used as a positive control. A depleted lysate with the GFP mRNA but without the test TREM added is used as a negative control. The progress of GFP mRNA translation is monitored by fluorescence increase on a microplate reader at 37° C. for 3-5 h using λ.sub.ex485/λ.sub.em528. It is expected for the experimental sample to be able to produce similar levels of fluorescence over time as the positive control and to be able to produce higher levels of fluorescence over time compared to the negative control. If so, these results would likely indicate that the test TREM is sufficient to, or can complement the depleted lysate and is thus likely functional.

Example 16: Assay for Modulation of Cell State

[1724] This example describes an assay for detecting activity of a TREM in modulating cell status, e.g., cell death.

[1725] TREM fragments are produced as described in Example 13. One (1) uM of TREM fragments are transfected into HEK293T cells with Lipofectamine 3000 and incubated for 1-6 hours in hour-long intervals followed by cell lysis. Cell lysates are analyzed by Western blotting and blots are probed with antibodies against total and cleaved caspase 3 and 9 as readouts of apoptosis. To measure cellular viability, cells are washed and fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature. Fixed and washed cells are then treated with 0.1% Triton X-100 for 10 minutes at room temperature and washed with PBS three times. Finally, cells are treated with TUNEL assay reaction mixture at 37° C. for 1 hour in the dark. Samples are analyzed by flow cytometry.

Example 17: Assay for the Activity of an Uncharged TREM to Modulate Autophagy

[1726] This example describes an assay to test an uncharged TREM for ability to modulate, e.g., induce, autophagy, e.g., the ability to activate GCN2-dependent stress response (starvation) pathway signaling, inhibit mTOR or activate autophagy.

[1727] A test uncharged TREM (uTREM) preparation is delivered to HEK293T or HeLa cells through transfection or liposomal delivery. Once the uTREM is delivered, a time course is performed ranging from 30 minutes to 6 hours with hour-long interval time points. Cells are then trypsinized, washed and lysed. The same procedure is executed with a charged control TREM as well as random RNA oligos as controls. Cell lysates are analyzed by Western blotting and blots are probed with antibodies against known readouts of GCN2 pathway activation, mTOR pathway inhibition or autophagy induction, including but not limited to phospho-eIF2a, ATF4, phospho-ULK1, phospho-4EBP1, phospho-eIF2a, phospho-Akt and phospho-p70S6K. A total protein loading control, such as GAPDH, actin or tubulin, as well as the non-modified (i.e. non-phosphorylated) signaling protein, i.e. using eIF2a as a control for phospho-eIF2a, are probed as loading controls. Delivery of the uTREM, compared to controls, is or can be expected to show activation of GCN2 starvation signaling pathway, autophagy pathway and/or inhibition of the mTOR pathway as determined by Western blot analysis.

Example 18: Assay for Activity of a Mischarged TREM (mTREM)

[1728] This example describes an assay to test the functionality of a mTREM produced in a cell system using plasmid transfection followed by in vitro mischarging.

[1729] In this example, an mTREM can translate a mutant mRNA into a wild type (WT) protein by incorporation of the WT amino acid in the protein despite an mRNA containing a mutated codon. GFP mRNA molecules with either a T203I or E222G mutation, which prevent GFP excitation at the 470 nm and 390 nm wavelengths, respectively, are used for this example. GFP mutants which prevent GFP fluorescence could also be used as reporter proteins in this assay. Briefly, an in vitro translation assay is used, using a rabbit reticulocyte lysate containing the GFP E222G mutated mRNA (GAG.fwdarw.GGG mutation) and an excess of the mTREM, in this case Glu-CCC-TREM. As a negative control, no mischarged TREM is added to the reaction. If the mTREM is functional, it is or can be expected that the GFP protein produced fluoresces when illuminated with a 390 nm excitation wavelength using a fluorimeter. If the mTREM is not functional or is less functional, the GFP protein produced fluoresces only when excited with a 470 nm wavelength, as is observed in the negative control.