RNA INTERFERENCE-INDUCING NUCLEIC ACID INHIBITING NONCANONICAL TARGETS OF MICRO RNA, AND USE FOR SAME
20220267766 · 2022-08-25
Assignee
Inventors
Cpc classification
C12N15/111
CHEMISTRY; METALLURGY
C12Q1/6876
CHEMISTRY; METALLURGY
C12N15/113
CHEMISTRY; METALLURGY
International classification
C12N15/113
CHEMISTRY; METALLURGY
Abstract
The present invention relates to RNA interference-inducing nucleic acid that inhibits noncanonical target genes of micro RNA, in which part of the sequence of a specific micro RNA has been modified, and by using the RNA interference-inducing nucleic acid of the present invention, the biological function micro RNA exhibits by inhibiting noncanonical target genes is effectively increased or there is the benefit of selectively exhibiting only one of the biological functions of conventional micro RNA, i.e., the function of inhibiting noncanonical target genes, and the interference-inducing nucleic acid of the present invention enables cell cycling, differentiation, dedifferentiation, formation, movement, splitting, proliferation or death adjustment, and it is expected that the invention can be used in various fields such as drugs and cosmetics.
Claims
1. An RNA interference-inducing nucleic acid, which suppresses a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of the nucleic acid inducing RNA interference, wherein the RNA interference-inducing nucleic acid has a sequence of four bases in positions 2 to 5 from the 5′ end of the specific miRNA, and bases in positions 6 and 7, which are the same and complementary to a base capable of pairing with the 6.sup.th base of the specific miRNA, including all complementary bases including G:A and G:U wobble pairs, to allow non-canonical target base pairs bound to a bulge generated in the target gene between positions 5 and 6 of the miRNA to be a consecutive base pair by the disappearance of the bulge, or the RNA interference-inducing nucleic acid has a modified base sequence in which at least one guanine is substituted with uracil or adenine in a base sequence between the first to ninth bases from the 5′ end of specific miRNA, in which a G:A or G:U wobble pair at the corresponding site becomes a canonical base sequence of U:A or A:U.
2. The RNA interference-inducing nucleic acid of claim 1, wherein the RNA interference-inducing nucleic acid selectively suppresses a non-canonical target gene of miRNA and does not suppress a canonical target gene of miRNA.
3. The RNA interference-inducing nucleic acid of claim 1, wherein the specific miRNA is one or more selected from the group consisting of miR-124, miR-155, miR-122, miR-1, let-7, miR-133, miR-302 and miR-372, which have the same seed sequence and consist of 18 to 24 bases.
4. The RNA interference-inducing nucleic acid of claim 1, wherein the RNA interference-inducing nucleic acid has the sequence from the 2n.sup.d to 7.sup.th bases from the 5′ end, which is one or more selected from: an RNA interference-inducing nucleic acid (miR-124-BS) having the base sequence of 5′-AA GGC C-3′, which is an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124; an RNA interference-inducing nucleic acid (miR-122-BS) having the base sequence of 5′-GG AGU U-3′, which is an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-122; an RNA interference-inducing nucleic acid (miR-155-BS) having the base sequence of 5′-UA AUG G-3′, which is an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-155; and an RNA interference-inducing nucleic acid (miR-1-BS) having the base sequence of 5′-GG AAU U-3′, which is an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-1.
5. The RNA interference-inducing nucleic acid of claim 4, wherein the RNA interference-inducing nucleic acid has any one or more base sequences as follows: an RNA interference-inducing nucleic acid (miR-124-BS) having a base sequence of 5′-UAA GGC CAC GCG GUG AAU GCC-3′ as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124; an RNA interference-inducing nucleic acid (miR-122-BS) having a base sequence of 5′-UGG AGU UGU GAC AAU GGU GUU-3′ as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-122; an RNA interference-inducing nucleic acid (miR-155-BS) having a base sequence of 5′-UUA AUG GC UAA U CGU GAU AGG-3′ as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-155; or an RNA interference-inducing nucleic acid (miR-1-BS) having a base sequence of 5′-UGG AAU UGU AAA GAA GUA UGU-3′ as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-1.
6. The RNA interference-inducing nucleic acid of claim 1, wherein the RNA interference-inducing nucleic acid has any one or more base sequences between the 1.sup.st to 9.sup.th bases from the 5′ end as follows: as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124, an RNA interference-inducing nucleic acid having a base sequence of 5′-UAA UGC ACG-3′ (miR-124-G4U), 5′-UAA GUC ACG-3′ (miR-124-G5U) or 5′-UAA UUC ACG-3′ (miR-124-G4,5U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-1, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUG AAU GUA-3′ (miR-1-G2U), 5′-UGU AAU GUA-3′ (miR-1-G3U), 5′-UGG AAU UUA-3′ (miR-1-G7U), 5′-UUU AAU GUA-3′ (miR-1-G2,3U), 5′-UGU AAU UUA-3′ (miR-1-G3,7U), 5′-UUG AAU UUA-3′ (miR-1-G2,7U) or 5′-UUU AAU UUA-3′ (miR-1-G2,3,7U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-122, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUG AGU GUG-3′ (miR-122-G2U), 5′-UGU AGU GUG-3′ (miR-122-G3U), 5′-UGG AUU GUG-3′ (miR-122-G5U), 5′-UGG AGU UUG-3′ (miR-122-G7U), 5′-UGG AGU GUU-3′ (miR-122-G9U), 5′-UUU AGU GUG-3′ (miR-122-G2,3U), 5′-UUG AUU GUG-3′ (miR-122-G2,5U), 5′-UUG AGU UUG-3′ (miR-122-G2,7U), 5′-UUG AGU GUU-3′ (miR-122-G2,9U), 5′-UGU AUU GUG (miR-122-G3,5U), 5′-UGU AGU UUG-3′ (miR-122-G3,7U), 5′-UGU AGU GUU-3′ (miR-122-G3,9U), 5′-UGG AUU UUG-3′ (miR-122-G5,7U), 5′-UGG AUU GUU-3′ (miR-122-G5,9U) or 5′-UGG AGU UUU-3 (miR-122-G7,9U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-133, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUU UGU CCC-3′ (miR-133-G4U), 5′-UUU GUU CCC-3′ (miR-133-G5U) or 5′-UUU UUU CCC-3′(miR-133-G4,5U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of let-7, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUA GGU AGU-3′ (let-7-G2U), 5′-UGA UGU AGU-3′ (let-7-G4U), 5′-UGA GUU AGU-3′ (let-7-G5U), 5′-UGA GGU AUU-3′ (let-7-G8U), 5′-UUA UGU AGU-3′ (let-7-G2,4U), 5′-UUA GUU AGU-3′ (let-7-G2,5U), 5′-UUA GGU AUU-3′ (let-7-G2,8U), 5′-UGA UUU AGU-3′ (let-7-G4,5U), 5′-UGA UGU AUU-3′ (let-7-G4,8U) or 5′-UGA GUU AUU-3′ (let-7-G5,8U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-302a, an RNA interference-inducing nucleic acid having a base sequence of 5′-UAA UUG CUU-3′ (miR-302a-G4U), 5′-UAA GUU CUU-3′ (miR-302a-G6U), or 5′-UAA UUU CUU-3′ (miR-302a-G4,6U); and as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-372, an RNA interference-inducing nucleic acid having a base sequence of 5′-AAA UUG CUG-3′ (miR-372-G4U), 5′-AAA GUU CUG-3′ (miR-372-G6U), 5′-AAA GUG CUU-3′ (miR-372-G9U), 5′-AAA UUU CUG-3′ (miR-372-G4,6U), 5′-AAA UUG CUU-3′ (miR-372-G4,9U) or 5′-AAA GUU CUU-3′ (miR-372-G6,9U).
7. The RNA interference-inducing nucleic acid of claim 6, wherein the RNA interference-inducing nucleic acid has one or more base sequences as follows: as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124, an RNA interference-inducing nucleic acid having a base sequence of 5′-UAA UGC ACG CGG UGA AUG CCA A-3′ (miR-124-G4U), 5′-UAA GUC ACG CGG UGA AUG CCA A-3′(miR-124-G5U) or 5′-UAA UUC ACG CGG UGA AUG CCA A-3′(miR-124-G4,5U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-1, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUG AAU GUA AAG AAG UAU GUA U-3′ (miR-1-G2U), 5′-UGU AAU GUA AAG AAG UAU GUA U-3′ (miR-1-G3U), 5′-UGG AAU UUA AAG AAG UAU GUA U-3′ (miR-1-G7U), 5′-UUU AAU GUA AAG AAG UAU GUA U-3′ (miR-1-G2,3U), 5′-UGU AAU UUA AAG AAG UAU GUA U-3′ (miR-1-G3,7U), 5′-UUG AAU UUA AAG AAG UAU GUA U-3′ (miR-1-G2,7U) or 5′-UUU AAU UUA AAG AAG UAU GUA U-3′ (miR-1-G2,3,7U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-122, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUG AGU GUG ACA AUG GUG UUU G-3′ (miR-122-G2U), 5′-UGU AGU GUG ACA AUG GUG UUU G-3 (miR-122-G3U), 5′-UGG AUU GUG ACA AUG GUG UUU G-3′ (miR-122-G5U), 5′-UGG AGU UUG ACA AUG GUG UUU G-3′ (miR-122-G7U), 5′-UGG AGU GUU ACA AUG GUG UUU G-3′ (miR-122-G9U), 5′-UUU AGU GUG ACA AUG GUG UUU G-3′ (miR-122-G2,3U), 5′-UUG AUU GUG ACA AUG GUG UUU G-3′ (miR-122-G2,5U), 5′-UUG AGU UUG ACA AUG GUG UUU G-3′ (miR-122-G2,7U), 5′-UUG AGU GUU ACA AUG GUG UUU G-3′ (miR-122-G2,9U), 5′-UGU AUU GUG ACA AUG GUG UUU G-3 (miR-122-G3,5U), 5′-UGU AGU UUG ACA AUG GUG UUU G-3 (miR-122-G3,7U), 5′-UGU AGU GUU ACA AUG GUG UUU G-3 (miR-122-G3,9U), 5′-UGG AUU UUG ACA AUG GUG UUU G-3′ (miR-122-G5,7U), 5′-UGG AUU GUU ACA AUG GUG UUU G-3′ (miR-122-G5,9U) or 5′-UGG AGU UUU ACA AUG GUG UUU G-3′ (miR-122-G7,9U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-133, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUU UGU CCC CUU CAA CCA GCU G -3′ (miR-133-G4U), 5′-UUU GUU CCC CUU CAA CCA GCU G-3′ (miR-133-G5U) or 5′-UUU UUU CCC CUU CAA CCA GCU G-3′(miR-133-G4,5U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of let-7, an RNA interference-inducing nucleic acid having a base sequence of 5′-UUA GGU AGU AGG UUG UAU AGU U-3′ (let-7-G2U), 5′-UGA UGU AGU AGG UUG UAU AGU U-3′ (let-7-G4U), 5′-UGA GUU AGU AGG UUG UAU AGU U-3′ (let-7-G5U), 5′-UGA GGU AUU AGG UUG UAU AGU U-3′ (let-7-G8U), 5′-UUA UGU AGU AGG UUG UAU AGU U-3′ (let-7-G2,4U), 5′-UUA GUU AGU AGG UUG UAU AGU U-3′ (let-7-G2,5U), 5′-UUA GGU AUU AGG UUG UAU AGU U-3′ (let-7-G2,8U), 5′-UGA UUU AGU AGG UUG UAU AGU U-3′ (let-7-G4,5U), 5′-UGA UGU AUU AGG UUG UAU AGU U-3′ (let-7-G4,8U) or 5′-UGA GUU AUU AGG UUG UAU AGU U-3′ (let-7-G5,8U); as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-302a, an RNA interference-inducing nucleic acid having a base sequence of 5′-UAA UUG CUU CCA UGU UUU GGU GA-3′ (miR-302a-G4U), 5′-UAA GUU CUU CCA UGU UUU GGU GA-3′ (miR-302a-G6U) or 5′-UAA UUU CUU CCA UGU UUU GGU GA-3′ (miR-302a-G4,6U); and as an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-372, an RNA interference-inducing nucleic acid having a base sequence of 5′-AAA UUG CUG CGA CAU UUG AGC GU -3′ (miR-372-G4U), 5′-AAA GUU CUG CGA CAU UUG AGC GU -3′ (miR-372-G6U), 5′-AAA GUG CUU CGA CAU UUG AGC GU -3′ (miR-372-G9U), 5′-AAA UUU CUG CGA CAU UUG AGC GU -3′ (miR-372-G4,6U), 5′-AAA UUG CUU CGA CAU UUG AGC GU -3′ (miR-372-G4,9U) or 5′-AAA GUU CUU CGA CAU UUG AGC GU -3′ (miR-372-G6,9U).
8. A composition for inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising the RNA interference-inducing nucleic acid of claim 1.
9. A composition for regulating cell cycling, differentiation, dedifferentiation, morphology, migration, division, proliferation or apoptosis, comprising the RNA interference-inducing nucleic acid of claim 1.
10. The composition of claim 9, wherein the composition is any one or more selected from: a composition for inducing cancer cell death, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124; a composition for inducing neurite or dendrite differentiation, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124; a composition for inducing cell cycle arrest in liver cancer cells, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-122; a composition for promoting differentiation of muscle cells or muscle fibrosis, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-1; a composition for inducing muscle cell death, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-155; a composition for inducing cell death of neuroblastomas, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124; a composition for promoting cell division or proliferation of neuroblastomas, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-124; a composition for inducing myocardial hypertrophy, which comprises an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-1; a composition for inducing myocardial hypertrophy, which includes an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-133; a composition for inducing cell cycle arrest in cancer cells, which includes an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of let-7; a composition for inducing the cell cycle progressing activity of hepatocytes, which includes an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of let-7; a composition for promoting dedifferentiation, which includes an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-302a; a composition for promoting dedifferentiation, which includes an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-372; and a composition for inhibiting cell migration of liver cancer cells, which includes an RNA interference-inducing nucleic acid suppressing a non-canonical target gene of miR-122.
11. A method of preparing an RNA interference-inducing nucleic acid inhibiting the expression of a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of the nucleic acid inducing RNA interference, the method comprising the following steps: constructing an RNA interference-inducing nucleic acid having a sequence of four bases in positions 2 to 5 from the 5′ end of specific miRNA and bases in positions 6 and 7, which are the same and complementary to a base capable of pairing with the 6.sup.th base of the specific miRNA, including all complementary bases including G:A and G:U wobble pairs, to allow non-canonical target base pairs bound to a bulge generated in the target gene between positions 5 and 6 of the miRNA to be a consecutive base pair by the disappearance of the bulge, or constructing an RNA interference-inducing nucleic acid having a modified base sequence in which at least one guanine is substituted with uracil or adenine in a base sequence between the first to ninth bases from the 5′ end of specific miRNA, in which a G:A or G:U wobble pair at the corresponding site becomes a canonical base sequence of U:A or A:U.
12. A method of screening a test material for regulating cell cycling, differentiation, dedifferentiation, morphology, migration, division, proliferation or apoptosis, comprising: transfecting the RNA interference-inducing nucleic acid of claim 1 into target cells; treating the target cells with a test material; and confirming an expression level or expression of a non-canonical target gene of microRNA (miRNA) inhibited by the RNA interference-inducing nucleic acid in the target cells.
13. An RNA interference-inducing nucleic acid suppressing a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of a nucleic acid inducing RNA interference, wherein the RNA interference-inducing nucleic acid has a sequence of four bases in positions 2 to 5 from the 5′ end of the specific miRNA, and bases in positions 6 and 7, which are the same and complementary to a base capable of pairing with the 6.sup.th base of specific miRNA, including all complementary bases including a G:A or G:U wobble pair, to allow non-canonical target base pairs bound to a bulge generated in the target gene between positions 5 and 6 of the miRNA to be a consecutive base pair by the disappearance of the bulge, and the specific miRNA has a methyl group (2′OMe) added to the 2′ position of the ribosyl ring of the 6.sup.th nucleotide from the 5′ end.
14. The RNA interference-inducing nucleic acid of claim 13, wherein the RNA interference-inducing nucleic acid inhibits only the expression of a canonical seed target gene of the corresponding RNA interference-inducing nucleic acid.
15. The RNA interference-inducing nucleic acid of claim 13, wherein the RNA interference-inducing nucleic acid specifically suppresses only a non-canonical nucleation bulge site of the specific miRNA, and removes the non-canonical nucleation bulge site which is able to be newly generated.
16. A composition for inhibiting the expression of a non-canonical target gene of microRNA (miRNA), which includes the RNA interference-inducing nucleic acid of claim 13.
17. A method of preparing an RNA interference-inducing nucleic acid, which inhibits the expression of a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of a nucleic acid inducing RNA interference, the method comprising the following steps: constructing an RNA interference-inducing nucleic acid having a sequence of four bases in positions 2 to 5 from the 5′ end of the specific miRNA, and bases in positions 6 and 7, which are the same and complementary to a base capable of pairing with the 6.sup.th base of the specific miRNA, including all complementary bases including G:A and G:U wobble pairs, to allow non-canonical target base pairs bound to a bulge generated in the target gene between positions 5 and 6 of the miRNA to be a consecutive base pair by the disappearance of the bulge; and adding a methyl group (2′OMe) to the 2′ position of the ribosyl ring of the 6.sup.th nucleotide from the 5′ end.
18. An RNA interference-inducing nucleic acid, which suppresses a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of a nucleic acid inducing RNA interference, wherein the RNA interference-inducing nucleic acid has a sequence of four bases in positions 2 to 5 from the 5′ end of the specific miRNA, and bases in positions 6 and 7, which are the same and complementary to a base capable of pairing with the 6.sup.th base of the specific miRNA, including all complementary bases including G:A and G:U wobble pairs, to allow non-canonical target base pairs bound to a bulge generated in the target gene between positions 5 and 6 of the miRNA to be a consecutive base pair by the disappearance of the bulge.
19. The RNA interference-inducing nucleic acid of claim 18, wherein the RNA interference-inducing nucleic acid selectively suppresses a non-canonical nucleation bulge target site and does not suppress a canonical target gene of miRNA.
20. The RNA interference-inducing nucleic acid of claim 18, wherein the RNA interference-inducing nucleic acid has a base sequence represented by any one of SEQ ID NOs: 103 to 528, shown in the following table: TABLE-US-00005 BS SEQ Bulge sequence ID NOs miRNA family Seed Seed site site GAGGUU 103 let-7/98/4458/4500 GAGGUA UACCUC AACCUC CCCUGG 104 miR-125a-5p/125b-5p/351/670/4319 CCCUGA UCAGGG CCAGGG AAGGCC 105 miR-124/124ab/506 AAGGCA UGCCUU GGCCUU CUUUGG 106 miR-9/9ab CUUUGG CCAAAG CCAAAG AGCACC 107 miR-29abcd AGCACC GGUGCU GGUGCU GCAGCC 108 miR-103a/107/107ab GCAGCA UGCUGC GGCUGC GCUACC 109 miR-221/222/222ab/1928 GCUACA UGUAGC GGUAGC UCAAGG 110 miR-26ab/1297/4465 UCAAGU ACUUGA CCUUGA AGCAGG 111 miR-15abc/16/16abc/195/322/424/497/1907 AGCAGC GCUGCU CCUGCU CGUACC 112 miR-126-3p CGUACC GGUACG GGUACG GUAAAA 113 mi R-30a bcdef/30a be-50/384-5p GUAAAC GUUUAC UUUUAC UGCAUU 114 miR-33ab/33-5p UGCAUU AAUGCA AAUGCA GGCAGG 115 miR-34ac/34bc-5p/449abc/449c-5p GGCAGU ACUGCC CCUGCC GUGCAA 116 miR-19ab GUGCAA UUGCAC UUGCAC ACCCGG 117 miR-99ab/100 ACCCGU ACGGGU CCGGGU AAAGUU 118 miR-17/17-5p/20ab/20b-5p/93/106ab/427/518a- AAAGUG CACUUU AACUUU 3p/519d UCACAA 119 miR-27abc/27a-3p UCACAG CUGUGA UUGUGA UGUGCC 120 miR-218/218a UGUGCU AGCACA GGCACA AGCUGG 121 miR-22/22-3p AGCUGC GCAGCU CCAGCU GGAGAA 122 mi R-185/882/3473/4306/4644 GGAGAG CUCUCC UUCUCC ACAUUU 123 miR-181abcd/4262 ACAUUC GAAUGU AAAUGU CCAGCC 124 miR-338/338-3p CCAGCA UGCUGG GGCUGG CGGAUU 125 miR-127/127-3p CGGAUC GAUCCG AAUCCG ACAGUU 126 miR-101/101ab ACAGUA UACUGU AACUGU CUGGCC 127 miR-149 CUGGCU AGCCAG GGCCAG GCAUCC 128 miR-324-5p GCAUCC GGAUGC GGAUGC GGCUCC 129 miR-24/24ab/24-3p GGCUCA UGAGCC GGAGCC AAUGCC 130 miR-33a-3p/365/365-3p AAUGCC GGCAUU GGCAUU CUACAA 131 miR-139-5p CUACAG CUGUAG UUGUAG GCUGGG 132 miR-138/138ab GCUGGU ACCAGC CCCAGC GAGAUU 133 miR-143/1721/4770 GAGAUG CAUCUC AAUCUC AUUGCC 134 miR-25/32/92abc/363/363-30/367 AUUGCA UGCAAU GGCAAU GAGUGG 135 miR-574-5p GAGUGU ACACUC CCACUC GGAAGG 136 miR-7/7ab GGAAGA UCUUCC CCUUCC UCCAGG 137 miR-145 UCCAGU ACUGGA CCUGGA AUGGCC 138 miR-135ab/135a-5p AUGGCU AGCCAU GGCCAU CAGUGG 139 miR-148ab-30/152 CAGUGC GCACUG CCACUG AGGAGG 140 miR-28-5 /708/1407/1653/3139 AGGAGC GCUCCU CCUCCU AGUGCC 141 miR-130ac/301ab/301b/301b-3p/454/721/4295/3666 AGUGCA UGCACU GGCACU GGGUAA 142 miR-3132 GGGUAG CUACCC UUACCC UAAUGG 143 miR-155 UAAUGC GCAUUA CCAUUA UCAUAA 144 miR-485-3p UCAUAC GUAUGA UUAUGA AACAGG 145 miR-132/212/212-3p AACAGU ACUGUU CCUGUU UAAAGG 146 hsa-m i R-9-3p UAAAGC GCUUUA CCUUUA UAUAAA 147 miR-374ab UAUAAU AUUAUA UUUAUA AGCCCC 148 miR-129-3p/129ab-3p/129-1-3p/129-2-3p AGCCCU AGGGCU GGGGCU AUUAUU 149 hsa-miR-126-5p AUUAUU AAUAAU AAUAAU AUGACC 150 miR-425/425-5 /489 AUGACA UGUCAU GGUCAU GCUCGG 151 miR-423-3p GCUCGG CCGAGC CCGAGC AGCUUU 152 miR-21/590-5p AGCUUA UAAGCU AAAGCU GGCAAA 153 miR-31 GGCAAG CUUGCC UUUGCC CUGUAA 154 hsa-miR-20b-3p CUGUAG CUACAG UUACAG UAUACC 155 hsa-let-7d-3p UAUACG CGUAUA GGUAUA AACGGG 156 miR-191 AACGGA UCCGUU CCCGUU AAGGUU 157 miR-18ab/4735-3p AAGGUG CACCUU AACCUU AUAAUU 158 miR-369-3p AUAAUA UAUUAU AAUUAU GGGAUU 159 hsa-miR-5187-5p GGGAUG CAUCCC AAUCCC AAGUUU 160 miR-382 AAGUUG CAACUU AAACUU GAGGCC 161 miR-485-5p/1698/1703/1962 GAGGCU AGCCUC GGCCUC AUCAUU 162 hsa-miR-136-3p AUCAUC GAUGAU AAUGAU AGAUGG 163 miR-576-3p AGAUGU ACAUCU CCAUCU UCCCUU 164 miR-204/204b/211 UCCCUU AAGGGA AAGGGA GAGACC 165 miR-769-5p GAGACC GGUCUC GGUCUC GGGGUU 166 miR-342-50/4664-5p GGGGUG CACCCC AACCCC UAUCAA 167 miR-361-5p UAUCAG CUGAUA UUGAUA CAGUAA 168 miR-199ab-30/3129-5p CAGUAG CUACUG UUACUG GUAGUU 169 miR-142-3p GUAGUG CACUAC AACUAC GGUUUU 170 miR-299-50/3563-5p GGUUUA UAAACC AAAACC ACUGGG 171 miR-193/193b/193a-3p ACUGGC GCCAGU CCCAGU AAUAUU 172 hsa-miR-1277-5p AAUAUA UAUAUU AAUAUU AGUGGG 173 miR-140/140-5p/876-3p/1244 AGUGGU ACCACU CCCACU UUUCAA 174 hsa-miR-30a/d/e-3p UUUCAG CUGAAA UUGAAA UGCGCC 175 hsa-let-7i-3p UGCGCA UGCGCA GGCGCA GGUUAA 176 miR-409-5 /409a GGUUAC GUAACC UUAACC GGUAGG 177 miR-379/1193-5 /3529 GGUAGA UCUACC CCUACC CUCCAA 178 miR-136 CUCCAU AUGGAG UUGGAG AGGUUU 179 miR-154/872 AGGUUA UAACCU AAACCU GUUGCC 180 miR-4684-3p GUUGCA UGCAAC GGCAAC CCCCCC 181 miR-361-3p CCCCCA UGGGGG GGGGGG CAAGAA 182 mi R-335/335-5p CAAGAG CUCUUG UUCUUG GAGGGG 183 miR-423a/423-5p/3184/3573-5p GAGGGG CCCCUC CCCCUC CUCAAA 184 miR-371/373/371b-5p CUCAAA UUUGAG UUUGAG GAGGAA 185 miR-1185/3679-5p GAGGAU AUCCUC UUCCUC CAAAAA 186 miR-3613-3p CAAAAA UUUUUG UUUUUG AAGUGG 187 miR-93/93a/105/106a/291a- AAGUGC GCACUU CCACUU 3p/294/295/302abcde/372/373/428/519a/520be/520 acd-30/1378/1420ac GGAUUU 188 miR-876-50/3167 GGAUUU AAAUCC AAAUCC ACACAA 189 miR-329/329ab/362-3p ACACAC GUGUGU UUGUGU UACAGG 190 miR-582-5p UACAGU ACUGUA CCUGUA GAGAAA 191 miR-146ac/146b-5p GAGAAC GUUCUC UUUCUC AUGUAA 192 miR-380/380-3p AUGUAA UUACAU UUACAU ACAUCC 193 miR-499-30/499a-3p ACAUCA UGAUGU GGAUGU CGACCC 194 miR-551a CGACCC GGGUCG GGGUCG AUAAAA 195 miR-142-5p AUAAAG CUUUAU UUUUAU CUGCAA 196 hsa-miR-17-3p CUGCAG CUGCAG UUGCAG CCAGUU 197 miR-199ab-5p CCAGUG CACUGG AACUGG GUGACC 198 miR-542-3p GUGACA UGUCAC GGUCAC ACGUAA 199 miR-1277 ACGUAG CUACGU UUACGU GACCGG 200 hsa-miR-29c-5p GACCGA UCGGUC CCGGUC GAUAUU 201 miR-3145-3p GAUAUU AAUAUC AAUAUC CGCACC 202 hsa-miR-106b-3p CGCACU AGUGCG GGUGCG GUUCUU 203 hsa-miR-22-5p GUUCUU AAGAAC AAGAAC GCGGGG 204 miR-744/1716 GCGGGG CCCCGC CCCCGC CCGUGG 205 hsa-miR-132-5p CCGUGG CCACGG CCACGG UGAAAA 206 mi R-488 UGAAAG CUUUCA UUUUCA AUGCAA 207 miR-501-3p/502-3p/500/502a AUGCAC GUGCAU UUGCAU CCUGUU 208 miR-486-5p/3107 CCUGUA UACAGG AACAGG UUUGCC 209 miR-450a/451a UUUGCG CGCAAA GGCAAA UGGGAA 210 hsa-miR-30c-3p UGGGAG CUCCCA UUCCCA UAAGAA 211 mi R-499-5p UAAGAC GUCUUA UUCUUA UCAACC 212 miR-421 UCAACA UGUUGA GGUUGA UCACCC 213 miR-197 UCACCA UGGUGA GGGUGA GGGCCC 214 miR-296-5p GGGCCC GGGCCC GGGCCC CUCUGG 215 miR-326/330/330-5p CUCUGG CCAGAG CCAGAG CAGCAA 216 miR-214/761/3619-5p CAGCAG CUGCUG UUGCUG CUGGGG 217 miR-612/1285/3187-5p CUGGGC GCCCAG CCCCAG AAUGUU 218 miR-409-3p AAUGUU AACAUU AACAUU CUGGAA 219 miR-378/422a/378bcdefh1 CUGGAC GUCCAG UUCCAG CUCACC 220 miR-342-3p CUCACA UGUGAG GGUGAG ACAAUU 221 miR-338-5p ACAAUA UAUUGU AAUUGU GGGGGG 222 miR-625 GGGGGA UCCCCC CCCCCC AAUACC 223 miR-200bc/429/548a AAUACU AGUAUU GGUAUU UAGAUU 224 hsa-miR-376a-5p UAGAUU AAUCUA AAUCUA UAUGGG 225 miR-584 UAUGGU ACCAUA CCCAUA AGUAGG 226 miR-411 AGUAGA UCUACU CCUACU UGAAGG 227 miR-573/3533/3616-50/3647-5p UGAAGU ACUUCA CCUUCA CCAUUU 228 miR-885-5p CCAUUA UAAUGG AAAUGG AAGCUU 229 hsa-miR-99-3p AAGCUC GAGCUU AAGCUU GGUGGG 230 miR-876-3p GGUGGU ACCACC CCCACC AUGUCC 231 miR-654-3p AUGUCU AGACAU GGACAU CCGUCC 232 hsa-miR-340-3p CCGUCU AGACGG GGACGG CACUUU 233 miR-3614-5p CACUUG CAAGUG AAAGUG GUGUUU 234 hsa-miR-124-5p GUGUUC GAACAC AAACAC GUGGGG 235 miR-491-5p GUGGGG CCCCAC CCCCAC UUGGCC 236 miR-96/507/1271 UUGGCA UGCCAA GGCCAA AAAACC 237 miR-548a-3p/548ef/2285a AAAACU AGUUUU GGUUUU AAUUUU 238 hsa-miR-32-3p AAUUUA UAAAUU AAAAUU AGCAAA 239 miR-3942-5p/4703-5p AGCAAU AUUGCU UUUGCU AGGCAA 240 miR-34b/449c/1360/2682 AGGCAG CUGCCU UUGCCU GGGUUU 241 hsa-miR-23a/b-5p GGGUUC GAACCC AAACCC AUCCUU 242 miR-362-5p/500b AUCCUU AAGGAU AAGGAU UCACUU 243 miR-677/4420 UCACUG CAGUGA AAGUGA AGAUAA 244 miR-577 AGAUAA UUAUCU UUAUCU GUUGUU 245 miR-3613-5p GUUGUA UACAAC AACAAC GAUCGG 246 miR-369-5p GAUCGA UCGAUC CCGAUC CUCCCC 247 miR-150/5127 CUCCCA UGGGAG GGGGAG UUCUGG 248 miR-544/544ab/544-3p UUCUGC GCAGAA CCAGAA CUGAUU 249 hsa-miR-29a-5p CUGAUU AAUCAG AAUCAG CAGGAA 250 miR-873 CAGGAA UUCCUG UUCCUG AGCCUU 251 miR-3614-3p AGCCUU AAGGCU AAGGCU AAAGAA 252 miR-186 AAAGAA UUCUUU UUCUUU CACUCC 253 miR-483-3p CACUCC GGAGUG GGAGUG UUAUCC 254 hsa-miR-374a-3p UUAUCA UGAUAA GGAUAA AGGUAA 255 miR-196abc AGGUAG CUACCU UUACCU GAUUCC 256 hsa-miR-145-3p GAUUCC GGAAUC GGAAUC UGGUUU 257 hsa-miR-29b-2-5p UGGUUU AAACCA AAACCA CCUGGG 258 hsa-miR-221-5p CCUGGC GCCAGG CCCAGG CCAAUU 259 miR-323b-3p CCAAUA UAUUGG AAUUGG GUCAUU 260 miR-616 GUCAUU AAUGAC AAUGAC CAAAGG 261 miR-330-3p CAAAGC GCUUUG CCUUUG AACAAA 262 hsa-miR-7-3p AACAAA UUUGUU UUUGUU CGUGUU 263 miR-187 CGUGUC GACACG AACACG CUAUUU 264 hsa-miR-26a-3p CUAUUC GAAUAG AAAUAG ACUGUU 265 miR-452/4676-3p ACUGUU AACAGU AACAGU UUUUUU 266 miR-129-5p/129ab-5p UUUUUG CAAAAA AAAAAA GUCAGG 267 miR-223 GUCAGU ACUGAC CCUGAC GCCAGG 268 miR-4755-3p GCCAGG CCUGGC CCUGGC CCCGUU 269 miR-1247 CCCGUC GACGGG AACGGG AACUAA 270 miR-3129-3p AACUAA UUAGUU UUAGUU UUUUCC 271 hsa-miR-335-3p UUUUCA UGAAAA GGAAAA CGGGGG 272 miR-542-5p CGGGGA UCCCCG CCCCCG CCAUCC 273 hsa-miR-181a-3p CCAUCG CGAUGG GGAUGG CCCAAA 274 hsa-miR-186-3p CCCAAA UUUGGG UUUGGG GAGCUU 275 hsa-miR-27b-5p GAGCUU AAGCUC AAGCUC UUAUGG 276 miR-491-3p UUAUGC GCAUAA CCAUAA GGCUGG 277 miR-4687-3p GGCUGU ACAGCC CCAGCC AGUUAA 278 hsa-miR-101-5p AGUUAU AUAACU UUAACU GAUCAA 279 miR-4772-5p GAUCAG CUGAUC UUGAUC UCCUAA 280 miR-337-3p UCCUAU AUAGGA UUAGGA GUGUAA 281 hsa-miR-223-5p GUGUAU AUACAC UUACAC CAAUAA 282 hsa-miR-16/195-3p CAAUAU AUAUUG UUAUUG UCGUGG 283 miR-3677-3p UCGUGG CCACGA CCACGA GGAGGG 284 hsa-miR-766-5p GGAGGA UCCUCC CCCUCC AUGUGG 285 miR-299/299-30/3563-3b AUGUGG CCACAU CCACAU GCUUUU 286 miR-3140-3p GCUUUU AAAAGC AAAAGC AUGCCC 287 miR-532-50/511 AUGCCU AGGCAU GGGCAU GCCUAA 288 hsa-miR-24-5p GCCUAC GUAGGC UUAGGC AUUCUU 289 miR-4778-5p AUUCUG CAGAAU AAGAAU GACACC 290 miR-642b GACACA UGUGUC GGUGUC AGACGG 291 miR-483-5p AGACGG CCGUCU CCGUCU GCACCC 292 miR-767-5p GCACCA UGGUGC GGGUGC GCUAUU 293 hsa-miR-31-3p GCUAUG CAUAGC AAUAGC ACGCUU 294 miR-574-3p ACGCUC GAGCGU AAGCGU AAGGAA 295 miR-3173-3p AAGGAG CUCCUU UUCCUU GGGAGG 296 miR-2127/4728-5p GGGAGG CCUCCC CCUCCC GCUUCC 297 hsa-miR-103a-2-5p GCUUCU AGAAGC GGAAGC AACACC 298 miR-3591-3p AACACC GGUGUU GGUGUU ACUAUU 299 hsa-miR-625-3p ACUAUA UAUAGU AAUAGU GAAUCC 300 hsa-miR-15b-3p GAAUCA UGAUUC GGAUUC AAAUGG 301 miR-522/518e/1422p AAAUGG CCAUUU CCAUUU AAAAAA 302 miR-548d-3 /548acbz AAAAAC GUUUUU UUUUUU UCAUCC 303 hsa-miR-452-3p UCAUCU AGAUGA GGAUGA UGACCC 304 miR-192/215 UGACCU AGGUCA GGGUCA UAGCAA 305 miR-1551/4524 UAGCAG CUGCUA UUGCUA UCGGGG 306 hsa-miR-425-3p UCGGGA UCCCGA CCCCGA AUCUGG 307 miR-3126-3p AUCUGG CCAGAU CCAGAU CACAAA 308 hsa-miR-125b-2-3p CACAAG CUUGUG UUUGUG CUGCCC 309 miR-324-3 /1913 CUGCCC GGGCAG GGGCAG AUCUUU 310 hsa-miR-141-5p AUCUUC GAAGAU AAAGAU GGGACC 311 hsa-miR-365a/b-5p GGGACU AGUCCC GGUCCC CUGGUU 312 hsa-miR-29b-1-5p CUGGUU AACCAG AACCAG GGUUGG 313 miR-563/380-5p GGUUGA UCAACC CCAACC UUGAGG 314 miR-1304 UUGAGG CCUCAA CCUCAA UCUCUU 315 miR-216c/1461/4684-5p UCUCUA UAGAGA AAGAGA UUUUAA 316 hsa-miR-2681-5p UUUUAC GUAAAA UUAAAA GUAACC 317 miR-194 GUAACA UGUUAC GGUUAC AGGGUU 318 miR-296-3p AGGGUU AACCCU AACCCU AUUUCC 319 hsa-miR-205-3p AUUUCA UGAAAU GGAAAU ACUCAA 320 miR-888 ACUCAA UUGAGU UUGAGU ACAUGG 321 miR-4802-3p ACAUGG CCAUGU CCAUGU UGUACC 322 hsa-let-7a/d-3b UGUACA UGUACA GGUACA GGGCUU 323 miR-762/4492/4498 GGGCUG CAGCCC AAGCCC UGUUGG 324 hsa-miR-744-3p UGUUGC GCAACA CCAACA AGUUCC 325 hsa-miR-148b-5p AGUUCU AGAACU GGAACU UUGACC 326 miR-514/514b-3p UUGACA UGUCAA GGUCAA ACUAGG 327 miR-28-3p ACUAGA UCUAGU CCUAGU GUGCCC 328 miR-550a GUGCCU AGGCAC GGGCAC CGGGUU 329 hsa-miR-125b-1-3p CGGGUU AACCCG AACCCG AUUCAA 330 hsa-miR-506-5p AUUCAG CUGAAU UUGAAU CACCUU 331 hsa-miR-1306-5p CACCUC GAGGUG AAGGUG CCUUGG 332 miR-3189-3p CCUUGG CCAAGG CCAAGG GGUGCC 333 miR-675-50/4466 GGUGCG CGCACC GGCACC AAUCAA 334 hsa-miR-34a-3p AAUCAG CUGAUU UUGAUU CCCUAA 335 hsa-miR-454-5p CCCUAU AUAGGG UUAGGG ACUGCC 336 miR-509-5p/509-3-5p/4418 ACUGCA UGCAGU GGCAGU GUUUUU 337 hsa-miR-19a/b-5p GUUUUG CAAAAC AAAAAC UUCCCC 338 miR-4755-5p UUCCCU AGGGAA GGGGAA CUGCUU 339 hsa-miR-93-3p CUGCUG CAGCAG AAGCAG ACCCAA 340 miR-3130-5 /4482 ACCCAG CUGGGU UUGGGU CCAGAA 341 hsa-m i R-488-5p CCAGAU AUCUGG UUCUGG UCCUGG 342 hsa-miR-378a-5p UCCUGA UCAGGA CCAGGA AGCCAA 343 miR-575/4676-5p AGCCAG CUGGCU UUGGCU CUCGGG 344 miR-1307 CUCGGC GCCGAG CCCGAG UUCAGG 345 miR-3942-3p UUCAGA UCUGAA CCUGAA UGUUCC 346 miR-4677-5p UGUUCU AGAACA GGAACA GAGCGG 347 miR-339-3p GAGCGC GCGCUC CCGCUC AAGAAA 348 mi R-548 b-3p AAGAAC GUUCUU UUUCUU GUUCCC 349 hsa-miR-642b-5p GUUCCC GGGAAC GGGAAC AUCCCC 350 miR-188-5p AUCCCU AGGGAU GGGGAU AACCCC 351 hsa-miR-652-5p AACCCU AGGGUU GGGGUU AGUCCC 352 miR-2114 AGUCCC GGGACU GGGACU GUGGCC 353 miR-3688-5p GUGGCA UGCCAC GGCCAC AGGCCC 354 hsa-miR-15a-3p AGGCCA UGGCCU GGGCCU ACCAUU 355 hsa-miR-181c-3p ACCAUC GAUGGU AAUGGU GGAGUU 356 miR-122/122a/1352 GGAGUG CACUCC AACUCC UAUUAA 357 miR-556-3p UAUUAC GUAAUA UUAAUA AUGGUU 358 hsa-miR-218-2-3p AUGGUU AACCAU AACCAU CUUGUU 359 miR-643 CUUGUA UACAAG AACAAG ACCACC 360 mi R-140-3p ACCACA UGUGGU GGUGGU AGUGAA 361 miR-1245 AGUGAU AUCACU UUCACU AUCAGG 362 hsa-miR-2115-3p AUCAGA UCUGAU CCUGAU AAAGCC 363 miR-518bcf/518a-30/518d-3p AAAGCG CGCUUU GGCUUU ACCUUU 364 miR-3200-3p ACCUUG CAAGGU AAAGGU CAACAA 365 mi R-545/3065/3065-5p CAACAA UUGUUG UUGUUG CUUCUU 366 miR-1903/4778-3p CUUCUU AAGAAG AAGAAG CUUAAA 367 hsa-miR-302a-5p CUUAAA UUUAAG UUUAAG UGAAUU 368 hsa-miR-183-3p UGAAUU AAUUCA AAUUCA GGGGAA 369 miR-3144-5p GGGGAC GUCCCC UUCCCC AACUGG 370 miR-582-3p AACUGG CCAGUU CCAGUU AAGAUU 371 miR-4662a-3p AAGAUA UAUCUU AAUCUU CCUGAA 372 miR-3140-5p CCUGAA UUCAGG UUCAGG UGCAAA 373 hsa-miR-106a-3p UGCAAU AUUGCA UUUGCA AUAGGG 374 hsa-miR-135a-3p AUAGGG CCCUAU CCCUAU CUGACC 375 miR-345/345-5p CUGACU AGUCAG GGUCAG CAGGUU 376 miR-125a-3 /1554 CAGGUG CACCUG AACCUG ACUCCC 377 miR-3145-5p ACUCCA UGGAGU GGGAGU UGUCCC 378 miR-676 UGUCCU AGGACA GGGACA GCCCUU 379 miR-3173-5p GCCCUG CAGGGC AAGGGC AGAGUU 380 hsa-miR-5586-3p AGAGUG CACUCU AACUCU CCGAGG 381 miR-615-3p CCGAGC GCUCGG CCUCGG AUGGAA 382 miR-3688-3p AUGGAA UUCCAU UUCCAU UAGCCC 383 miR-4662a-5p UAGCCA UGGCUA GGGCUA UGCCAA 384 miR-4659ab-5p UGCCAU AUGGCA UUGGCA AUCCAA 385 hsa-miR-5586-5p AUCCAG CUGGAU UUGGAU ACUCUU 386 hsa-miR-514a-5p ACUCUG CAGAGU AAGAGU ACCCUU 387 miR-10abc/10a-5p ACCCUG CAGGGU AAGGGU ACUGAA 388 hsa-m i R-888-3p ACUGAC GUCAGU UUCAGU UCAGGG 389 miR-3127-5p UCAGGG CCCUGA CCCUGA GAUUGG 390 miR-508-3p GAUUGU ACAAUC CCAAUC GGGGCC 391 hsa-miR-185-3p GGGGCU AGCCCC GGCCCC GUCUUU 392 hsa-miR-200c-5p,hsa-miR-550a-3p GUCUUA UAAGAC AAAGAC UCUCAA 393 miR-513c/514b-5p UCUCAA UUGAGA UUGAGA AACCUU 394 miR-490-3p AACCUG CAGGUU AAGGUU CUGAAA 395 hsa-miR-5187-3p CUGAAU AUUCAG UUUCAG CUCAGG 396 miR-3664-3p CUCAGG CCUGAG CCUGAG GCCCCC 397 miR-3189-5p GCCCCA UGGGGC GGGGGC GAAGUU 398 miR-4670-3p GAAGUU AACUUC AACUUC CAAAUU 399 miR-105/105ab CAAAUG CAUUUG AAUUUG UGUAGG 400 hsa-miR-135b-3p UGUAGG CCUACA CCUACA UUUGUU 401 hsa-miR-5010-3p UUUGUG CACAAA AACAAA GAAGGG 402 miR-493/493b GAAGGU ACCUUC CCCUUC CUCCGG 403 miR-3605-3p CUCCGU ACGGAG CCGGAG UCCCAA 404 miR-188-3p UCCCAC GUGGGA UUGGGA UGCUAA 405 hsa-miR-449c-3p UGCUAG CUAGCA UUAGCA CAAGGG 406 miR-4761-5p CAAGGU ACCUUG CCCUUG AAGUCC 407 miR-224 AAGUCA UGACUU GGACUU GUCUAA 408 miR-4796-5p GUCUAU AUAGAC UUAGAC AAAUCC 409 hsa-miR-551b-5p AAAUCA UGAUUU GGAUUU AUGAGG 410 miR-556-5p AUGAGC GCUCAU CCUCAU ACGCCC 411 hsa-miR-122-3p ACGCCA UGGCGU GGGCGU CUGUGG 412 miR-4677-3p CUGUGA UCACAG CCACAG UAGAGG 413 miR-877 UAGAGG CCUCUA CCUCUA UUCUAA 414 miR-576-5p UUCUAA UUAGAA UUAGAA CAUGGG 415 miR-490-5p CAUGGA UCCAUG CCCAUG CAGAAA 416 hsa-miR-589-3p CAGAAC GUUCUG UUUCUG GAAGCC 417 miR-4786-3p GAAGCC GGCUUC GGCUUC UUAGCC 418 hsa-miR-374b-3p UUAGCA UGCUAA GGCUAA CUGUUU 419 hsa-miR-26b-3p CUGUUC GAACAG AAACAG AGGGCC 420 miR-3158-3p AGGGCU AGCCCU GGCCCU UAGGCC 421 miR-4423-3p UAGGCA UGCCUA GGCCUA UCUAGG 422 miR-518d-5p/519bc-5p520c-5p/523b/526a UCUAGA UCUAGA CCUAGA GCCCGG 423 miR-4707-3p GCCCGC GCGGGC CCGGGC AAAUUU 424 hsa-miR-10a-3p AAAUUC GAAUUU AAAUUU UCUUGG 425 miR-526b UCUUGA UCAAGA CCAAGA CUUCAA 426 hsa-miR-676-5p CUUCAA UUGAAG UUGAAG CCUCCC 427 hsa-miR-660-3p CCUCCU AGGAGG GGGAGG UUGGAA 428 hsa-miR-5004-3p UUGGAU AUCCAA UUCCAA GGGUCC 429 miR-193a-5p GGGUCU AGACCC GGACCC UCAGUU 430 hsa-miR-222-5p UCAGUA UACUGA AACUGA AGGAUU 431 miR-4661-3p AGGAUC GAUCCU AAUCCU GGCGGG 432 hsa-miR-25-5p GGCGGA UCCGCC CCCGCC AGCGAA 433 miR-4670-5p AGCGAC GUCGCU UUCGCU UUGGUU 434 miR-659 UUGGUU AACCAA AACCAA GCUCUU 435 miR-1745/3194-3p GCUCUG CAGAGC AAGAGC GGUUCC 436 hsa-miR-182-3p GGUUCU AGAACC GGAACC GCAGAA 437 miR-298/2347/2467-3p GCAGAG CUCUGC UUCUGC CUCUUU 438 hsa-miR-130b-5p CUCUUU AAAGAG AAAGAG GCGGUU 439 miR-4746-3p GCGGUG CACCGC AACCGC GCGCGG 440 miR-1893/2277-5p GCGCGG CCGCGC CCGCGC GGACCC 441 miR-3619-3p GGACCA UGGUCC GGGUCC CUACUU 442 hsa-miR-138-1-3p CUACUU AAGUAG AAGUAG AUGCUU 443 miR-4728-3p AUGCUG CAGCAU AAGCAU CCCCUU 444 miR-3127-3p CCCCUU AAGGGG AAGGGG CCGGUU 445 miR-671-3p CCGGUU AACCGG AACCGG CAGGGG 446 hsa-miR-211-3p CAGGGA UCCCUG CCCCUG GAGCCC 447 hsa-miR-2114-3p GAGCCU AGGCUC GGGCUC CCUCUU 448 hsa-miR-877-3p CCUCUU AAGAGG AAGAGG UCAGCC 449 miR-3157-5p UCAGCC GGCUGA GGCUGA UCCUUU 450 miR-502-5p UCCUUG CAAGGA AAAGGA AAUCCC 451 miR-500a AAUCCU AGGAUU GGGAUU AAACUU 452 miR-548Q AAACUG CAGUUU AAGUUU AACGCC 453 miR-523 AACGCG CGCGUU GGCGUU CAGUUU 454 hsa-miR-584-3p CAGUUC GAACUG AAACUG CCUUCC 455 miR-205/205ab CCUUCA UGAAGG GGAAGG CAUCCC 456 miR-4793-5p CAUCCU AGGAUG GGGAUG GGGUGG 457 hsa-miR-363-5p GGGUGG CCACCC CCACCC GCCUGG 458 hsa-miR-214-5p GCCUGU ACAGGC CCAGGC UUCCAA 459 miR-3180-5p UUCCAG CUGGAA UUGGAA UGGGGG 460 miR-1404/2110 UGGGGA UCCCCA CCCCCA UGCCCC 461 miR-3157-3p UGCCCU AGGGCA GGGGCA CUGCGG 462 hsa-miR-191-3p CUGCGC GCGCAG CCGCAG UGGGUU 463 miR-1346/3940-50/4507 UGGGUU AACCCA AACCCA CGGUCC 464 miR-4746-5p CGGUCC GGACCG GGACCG ACGCGG 465 miR-3939 ACGCGC GCGCGU CCGCGU CCACUU 466 hsa-miR-181a-2-3p CCACUG CAGUGG AAGUGG UGCACC 467 hsa-miR-500a-3p UGCACC GGUGCA GGUGCA CGACAA 468 hsa-m i R-196 b-3p CGACAG CUGUCG UUGUCG UGUAUU 469 hsa-miR-675-3p UGUAUG CAUACA AAUACA GCAAAA 470 hsa-miR-548a1/Q/x-5p GCAAAA UUUUGC UUUUGC UUCUUU 471 miR-4659ab-3p UUCUUC GAAGAA AAAGAA UCUGCC 472 hsa-miR-5001-3p UCUGCC GGCAGA GGCAGA CCCGGG 473 hsa-miR-1247-3p CCCGGG CCCGGG CCCGGG CCCCGG 474 miR-2890/4707-5p CCCCGG CCGGGG CCGGGG UGGUAA 475 hsa-miR-150-3p UGGUAC GUACCA UUACCA UUCUCC 476 hsa-miR-629-3p UUCUCC GGAGAA GGAGAA GACAGG 477 miR-2277-3p GACAGC GCUGUC CCUGUC GAGCAA 478 miR-3547/3663-3p GAGCAC GUGCUC UUGCUC AUCACC 479 miR-34bc-3p AUCACU AGUGAU GGUGAU AAGCGG 480 miR-518ef AAGCGC GCGCUU CCGCUU UGGCCC 481 miR-3187-3p UGGCCA UGGCCA GGGCCA CGUUGG 482 miR-1306/1306-3p CGUUGG CCAACG CCAACG GCACGG 483 miR-3177-3p GCACGG CCGUGC CCGUGC GGAAUU 484 miR-1ab/206/613 GGAAUG CAUUCC AAUUCC CACAGG 485 miR-128/128ab CACAGU ACUGUG CCUGUG UAGGGG 486 miR-1296 UAGGGC GCCCUA CCCCUA ACGUCC 487 miR-598/598-3p ACGUCA UGACGU GGACGU UGAACC 488 miR-887 UGAACG CGUUCA GGUUCA CAUACC 489 miR-1-5p CAUACU AGUAUG GGUAUG ACAUAA 490 miR-376c/741-5p ACAUAG CUAUGU UUAUGU UAAUAA 491 miR-374c/655 UAAUAC GUAUUA UUAUUA GAAACC 492 mi R-494 GAAACA UGUUUC GGUUUC UUAGGG 493 miR-651 UUAGGA UCCUAA CCCUAA UGCAGG 494 miR-1301/5047 UGCAGC GCUGCA CCUGCA GCGAGG 495 miR-381-5p GCGAGG CCUCGC CCUCGC AAUCUU 496 miR-216a AAUCUC GAGAUU AAGAUU AUACAA 497 mi R-300/381/539-3p AUACAA UUGUAU UUGUAU CGCCCC 498 miR-1249 CGCCCU AGGGCG GGGGCG UCAUUU 499 miR-579 UCAUUU AAAUGA AAAUGA AUAUUU 500 miR-656 AUAUUA UAAUAU AAAUAU UCAUGG 501 miR-433 UCAUGA UCAUGA CCAUGA UUCCGG 502 miR-1180 UUCCGG CCGGAA CCGGAA GUGUCC 503 miR-597/1970 GUGUCA UGACAC GGACAC UAUAUU 504 miR-190a-3p UAUAUA UAUAUA AAUAUA AAACCC 505 miR-1537 AAACCG CGGUUU GGGUUU GGCCCC 506 miR-874-5p GGCCCC GGGGCC GGGGCC AUAUAA 507 miR-410/344de/344b-1-3p AUAUAA UUAUAU UUAUAU CCUGCC 508 miR-370 CCUGCU AGCAGG GGCAGG GAAUUU 509 miR-219-2-3p/219-3p GAAUUG CAAUUC AAAUUC CACCCC 510 miR-3620 CACCCU AGGGUG GGGGUG GACCCC 511 miR-504/4725-5p GACCCU AGGGUC GGGGUC GAUGUU 512 miR-2964/2964a-5p GAUGUC GACAUC AACAUC UUGGGG 513 miR-450a-2-3p UUGGGG CCCCAA CCCCAA UGUCUU 514 miR-511 UGUCUU AAGACA AAGACA GACUUU 515 miR-6505-3p GACUUC GAAGUC AAAGUC ACGGUU 516 miR-433-5p ACGGUG CACCGU AACCGU CGGCUU 517 miR-6741-3p CGGCUC GAGCCG AAGCCG AGGUCC 518 miR-370-5p AGGUCA UGACCU GGACCU CGCGGG 519 miR-579-5p CGCGGU ACCGCG CCCGCG GUGGAA 520 miR-376c-5p,miR-376b-5p GUGGAU AUCCAC UUCCAC ACAGGG 521 miR-552/3097-5p ACAGGU ACCUGU CCCUGU CAGUCC 522 miR-1910 CAGUCC GGACUG GGACUG UUGUGG 523 miR-758 UUGUGA UCACAA CCACAA GGCCUU 524 miR-6735-3p GGCCUG CAGGCC AAGGCC GUAGAA 525 miR-376a-2-5p GUAGAU AUCUAC UUCUAC GGGCGG 526 miR-585 GGGCGU ACGCCC CCGCCC AACCGG 527 miR-451 AACCGU ACGGUU CCGGUU UAUUGG 528 miR-137/137ab UAUUGC GCAAUA CCAAUA
21. A composition for inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising the RNA interference-inducing nucleic acid of claim 18.
22. A method of preparing an RNA interference-inducing nucleic acid, which inhibits the expression of a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of a nucleic acid inducing RNA interference, the method comprising the following steps: constructing an RNA interference-inducing nucleic acid having a sequence of four bases in positions 2 to 5 from the 5′ end of the specific miRNA, and bases in positions 6 and 7, which are the same and complementary to a base capable of pairing with the 6.sup.th base of the specific miRNA, including all complementary bases including G:A and G:U wobble pairs, to allow non-canonical target base pairs bound to a bulge generated in the target gene between positions 5 and 6 of the miRNA to be a consecutive base pair by the disappearance of the bulge.
23. An RNA interference-inducing nucleic acid suppressing a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of a nucleic acid inducing RNA interference, wherein the RNA interference-inducing nucleic acid has a modified base sequence in which at least one guanine is substituted with uracil in a base sequence between the second to ninth bases from the 5′ end of specific miRNA, and the G:A wobble at the corresponding site becomes the canonical base pair of U:A.
24. The RNA interference-inducing nucleic acid of claim 23, wherein the RNA interference-inducing nucleic acid has a sequence of 6 to 8 consecutive bases, starting from the 2.sup.nd base from the 5′ end of specific miRNA, and the base sequence has at least one guanine base substituted with an uracil base.
25. The RNA interference-inducing nucleic acid of claim 23, wherein the RNA interference-inducing nucleic acid selectively suppresses only a non-canonical target gene of miRNA binding in a G:A wobble pair, and does not suppress a canonical target gene of miRNA.
26. The RNA interference-inducing nucleic acid of claim 24, wherein the RNA interference-inducing nucleic acid has the sequence of 6 to 8 consecutive bases, starting from the 2.sup.nd base from the 5′ end, represented by any one or more of SEQ ID NOs: 529 to 863 shown in the following table. TABLE-US-00006 G > T Sequence SEQ read/ (G > U) ID NOs miRNA name Seed Position patient UGAAUG 529 hsa-miR-1-3p GGAAUG 2 17.1 UUAACA 530 hsa-miR-194-5p GUAACA 2 96.3 UGGUCU 531 hsa-miR-193a-5p GGGUCU 2 12 UAAUCA 532 hsa-miR-15b-3p GAAUCA 2 2.2 UUCUUA 533 hsa-miR-200c-5p GUCUUA 2 4.2 UCCUGU 534 hsa-miR-214-5p GCCUGU 2 2.8 UUGACU 535 hsa-miR-134-5p GUGACU 2 10.3 UAUUCC 536 hsa-miR-145-3p GAUUCC 2 2.2 UUUCUU 537 hsa-miR-22-5p GUUCUU 2 2.5 UCUCGG 538 hsa-miR-423-3p GCUCGG 2 6.3 UAGACU 539 hsa-miR-873-3p GAGACU 2 4.5 UGAGUG 540 hsa-miR-122-5p GGAGUG 2 837.8 UAGAUG 541 hsa-miR-143-3p GAGAUG 2 322.7 UAGGCU 542 hsa-miR-485-5p GAGGCU 2 2.2 UGUUAC 543 hsa-miR-409-5p GGUUAC 2 8.1 UGCUCA 544 hsa-miR-24-3p GGCUCA 2 65.6 UUCAGU 545 hsa-miR-223-3p GUCAGU 2 12.7 UAUAUC 546 hsa-miR-144-5p GAUAUC 2 16.4 UGUAGA 547 hsa-miR-379-5p GGUAGA 2 47.2 UAGAAC 548 hsa-miR-146b-5p/hsa-miR-146a-5p GAGAAC 2 21.2 UAGAAA 549 hsa-miR-539-5p GAGAAA 2 3.3 UGGCCC 550 hsa-miR-296-5p GGGCCC 2 2.7 UCACCA 551 hsa-miR-767-5p GCACCA 2 10.1 UGCAGU 552 hsa-miR-34a-5p/hsa-miR-34c-5p GGCAGU 2 8.9 UAGGUA 553 hsa-let-7f-5p/hsa-let-7d-5p/hsa-let-7b-5p/hsa-let-7a- GAGGUA 2 320.1 5p/hsa-let-7e-5p/hsa-miR-202-3p/hsa-let-71-5p/hsa- miR-98-5 hsa-let-7c-5 /hsa-let-7 -5 UUGCCU 554 hsa-miR-1271-3p GUGCCU UCUGGU 555 hsa-miR-138-5p GCUGGU 2 5.9 UUGCAA 556 hsa-miR-19b-3p/hsa-miR-19a-3p GUGCAA 2 4.1 UGGCUU 557 hsa-miR-27a-5p GGGCUU 2 2.2 UCCCUG 558 hsa-miR-146b-3p GCCCUG 2 7.6 UGAAGA 559 hsa-miR-7-5p GGAAGA 2 2.3 UAGGGG 560 hsa-miR-423-5p GAGGGG 2 3.7 UCAUCC 561 hsa-miR-324-5p GCAUCC 2 2.6 UGGUUU 562 hsa-miR-629-5p GGGUUU 2 3.3 UGAGAC 563 hsa-miR-139-3p GGAGAC 2 2.3 UUAAAC 564 hsa-miR-30d-5p/hsa-miR-30e-5p/hsa-miR-30a- GUAAAC 2 1004.2 5p/hsa-miR-30c-5/hsa-miR-30b-5 UCUACA 565 hsa-miR-221-3p/hsa-miR-222-3p GCUACA 2 6.3 UAUUGG 566 hsa-miR-509-3p GAUUGG 2 54 UAGACC 567 hsa-miR-769-5p GAGACC 2 3.9 UUAGUG 568 hsa-miR-142-3p GUAGUG 2 5.2 UGAGAG 569 hsa-miR-185-5p GGAGAG 2 8.9 UAUUGU 570 hsa-miR-508-3p/hsa-miR-219a-5p GAUUGU 2 211.9 UGCAAG 571 hsa-miR-31-5p GGCAAG 2 4.2 UCAGCA 572 hsa-miR-103a-3p/hsa-miR-107 GCAGCA 2 757.3 UUGACA 573 hsa-miR-542-3p GUGACA 2 7.3 UAAUUG 574 hsa-miR-219a-2-3p GAAUUG 2 29.7 AUCACC 575 hsa-miR-29c-3p/hsa-miR-29a-3p/hsa-miR-29b-3p AGCACC 3 11 CUGGUU 576 hsa-miR-125b-1-3p CGGGUU 3 8.3 GUAAGA 577 hsa-miR-7-5p GGAAGA 3 2.3 AUUAGA 578 hsa-miR-411-5p AGUAGA 3 2.7 AUGUAG 579 hsa-miR-196a-5p/hsa-miR-196b-5p AGGUAG 3 3.3 AUGCAC 580 hsa-miR-3622a-5p AGGCAC 3 2 UUAAGC 581 hsa-miR-127-5p UGAAGC 3 4.6 AUCUGC 582 hsa-miR-22-3p AGCUGC 3 335.7 UUCAUA 583 hsa-miR-153-3p UGCAUA 3 3.1 GUGUCU 584 hsa-miR-193a-5p GGGUCU 3 5.8 GUAGAG 585 hsa-miR-185-5p GGAGAG 3 3i GUAAUG 586 hsa-miR-1-3p GGAAUG 3 10.2 AUCAGC 587 hsa-miR-15b-5p/hsa-miR-16-5p/hsa-miR-424-5p AGCAGC 3 5.6 UUUACA 588 hsa-let-7 -3 /hsa-miR-493-5 /hsa-let-7c-3 UGUACA 3 2.7 UUCGCA 589 hsa-let-7i-3p UGCGCA 3 2.4 UUUGCU 590 hsa-miR-218-5p UGUGCU 3 3.6 CUACCG 591 hsa-miR-1307-5p CGACCG 3 3.3 CUGAUC 592 hsa-miR-127-3p CGGAUC 3 10.5 UUUGCG 593 hsa-miR-210-3p UGUGCG 3 13.6 CUUGUC 594 hsa-miR-187-3p CGUGUC 3 2.1 UUCCAA 595 hsa-miR-192-3p UGCCAA 3 2.2 UUACCU 596 hsa-miR-192-5p UGACCU 3 73.2 GUCAGU 597 hsa-miR-34a-5p/hsa-miR-34c-5p GGCAGU 3 3.1 AUCUUA 598 hsa-miR-21-5p AGCUUA 3 738.6 UUCACC 599 hsa-miR-500a-3p UGCACC 3 2.7 GUGUUU 600 hsa-miR-629-5p GGGUUU 3 2.9 GUUAGA 601 hsa-miR-379-5p GGUAGA 3 22.1 UUAAAU 602 hsa-miR-203a-3p UGAAAU 3 200.1 GUCUCA 603 hsa-miR-24-3p GGCUCA 3 10.5 UUGGAG 604 hsa-miR-30c-2-3p UGGGAG 3 6.2 UUAAAG 605 hsa-miR-488-3p UGAAAG 3 2 AUUGCA 606 hsa-miR-301a-3p/hsa-miR-301b-3p AGUGCA 3 5 CUUACC 607 hsa-miR-126-3p CGUACC 3 15.8 GUAGUG 608 hsa-miR-122-5p GGAGUG 3 60.4 GUGCUU 609 hsa-miR-27a-5p GGGCUU 3 2.8 CUUUUC 610 hsa-miR-26b-3p CUGUUC 4 3 CUUCCC 611 hsa-miR-324-3p CUGCCC 4 2.3 CAUCAC 612 hsa-miR-3065-3p CAGCAC 4 4 GAUGGG 613 hsa-miR-423-5p GAGGGG 4 11.4 GUUUUC 614 hsa-miR-124-5p GUGUUC 4 3.3 CUUACU 615 hsa-miR-345-5p CUGACU 4 6 CCUAGC 616 hsa-miR-615-3p CCGAGC 4 2 AUUGCU 617 hsa-miR-889-5p/hsa-miR-135a-5p/hsa-miR-135b-5p AUGGCU 4 15.8 GGUUCU 618 hsa-miR-193a-5p GGGUCU 4 35.4 AAUGUG 619 hsa-miR-18a-5p AAGGUG 4 3.3 CGUGUU 620 hsa-miR-125b-1-3p CGGGUU 4 57 AGUAGC 621 hsa-miR-708-5p/hsa-miR-28-5p AGGAGC 4 10.5 AAUUCA 622 hsa-miR-224-5p AAGUCA 4 2.9 AAUCUU 623 hsa-miR-100-3p AAGCUU 4 9.5 CAUGAA 624 hsa-miR-873-5p CAGGAA 4 6.7 UAUCCA 625 hsa-miR-4662a-5p UAGCCA 4 7.5 AAUCUC 626 hsa-miR-99b-3p/hsa-miR-99a-3p AAGCUC 4 9.3 ACUGUG 627 hsa-miR-433-5p ACGGUG 4 2.3 GUUACA 628 hsa-miR-542-3p GUGACA 4 20.9 GAUGAU 629 hsa-miR-3605-5p GAGGAU 4 11.8 GCUGGG 630 hsa-miR-744-5p GCGGGG 4 4.2 UAUGGC 631 hsa-miR-1296-5p UAGGGC 4 26.2 UUUGUC 632 hsa-miR-133a-3p UUGGUC 4 11.7 AAUUUG 633 hsa-miR-382-5p AAGUUG 4 6.8 AUUACA 634 hsa-miR-425-5p AUGACA 4 2a9 GAUGUU 635 hsa-miR-377-5p GAGGUU 4 2.2 CGUAUC 636 hsa-miR-127-3p CGGAUC 4 219.1 GGUGCG 637 hsa-miR-3180-3p GGGGCG 4 2.4 GAUAUG 638 hsa-miR-143-3p GAGAUG 4 936.3 UUUUGA 639 hsa-miR-758-3p UUGUGA 4 4.9 CUUCUG 640 hsa-miR-93-3p CUGCUG 4 2.9 GGUGCC 641 hsa-miR-128-2-5p GGGGCC 4 4.2 GUUACU 642 hsa-miR-134-5p GUGACU 4 29.2 AGUUUA 643 hsa-miR-154-5p AGGUUA 4 3.5 AGUCAC 644 hsa-miR-3622a-5p AGGCAC 4 5.1 AAUGCA 645 hsa-miR-124-3p AAGGCA 4 38.7 GGUCUU 646 hsa-miR-27a-5p GGGCUU 4 5 CAUUGG 647 hsa-miR-194-3p CAGUGG 4 8.7 UUUUUC 648 hsa-miR-375 UUGUUC 4 1084.1 AAUUUC 649 hsa-miR-148a-5p AAGUUC 4 4.4 GCUCGG 650 hsa-miR-2277-5p GCGCGG 4 2.9 GAUACC 651 hsa-miR-769-5p GAGACC 4 26.3 CUUCAG 652 hsa-miR-17-3p CUGCAG 4 52.2 GAUACU 653 hsa-miR-873-3p GAGACU 4 9.9 CUUCAA 654 hsa-miR-4772-3p CUGCAA 4 2.3 AGUUUU 655 hsa-miR-329-5p AGGUUU 4 2.1 UUUGCA 656 hsa-miR-182-5p/hsa-miR-96-5p UUGGCA 4 598.4 GAUGCU 657 hsa-miR-2467-5p/hsa-miR-485-5p GAGGCU 4 12.7 CUUGCU 658 hsa-miR-149-5p CUGGCU 4 11.8 UGUUUU 659 hsa-miR-29b-2-5p UGGUUU 4 4.8 ACUCCA 660 hsa-miR-122-3p ACGCCA 4 5 AAUUGC 661 hsa-miR-302a-3p/hsa-miR-520a-3p/hsa-miR-519b- AAGUGC 4 29.3 3p/hsa-miR-520b/hsa-miR-519c-3p/hsa-miR-520c- 2p/hsa-miR-519a-3p AUUCCU 662 hsa-miR-532-5p AUGCCU 4 194.9 CCUUGG 663 hsa-miR-132-5p CCGUGG 4 2.3 AAUGAU 664 hsa-miR-541-5p AAGGAU 4 2.8 CCUGUU 665 hsa-miR-671-3p CCGGUU 4 3.6 GGUCCC 666 hsa-miR-296-5p GGGCCC 4 3 AAUCGC 667 hsa-miR-518e-3p AAGCGC 4 10.8 UGUUUA 668 hsa-miR-487a-5p UGGUUA 4 2.5 GAUAAC 669 hsa-miR-589-5p/hsa-miR-146b-5p/hsa-miR-146a-5p GAGAAC 4 60.3 AGUUAG 670 hsa-miR-196b-5p/hsa-miR-196a-5p AGGUAG 4 34.4 GGUGCA 671 hsa-miR-486-3p GGGGCA 4 2 GGUUUU 672 hsa-miR-629-5p GGGUUU 4 8.7 CUUGAC 673 hsa-miR-378a-3p CUGGAC 4 105.2 GAUCUU 674 hsa-miR-27b-5p GAGCUU 4 3.2 GCUCCU 675 hsa-miR-6720-3p GCGCCU 4 2.3 ACUCUC 676 hsa-miR-574-3p ACGCUC 4 16.6 CUUAUU 677 hsa-miR-29a-5p CUGAUU 4 2.6 UGUGAG 678 hsa-miR-30c-2-3p/hsa-miR-30c-1-3p UGGGAG 4 22.2 CAUUAG 679 hsa-miR-199b-3p CAGUAG 4 32.1 GAUUGU 680 hsa-miR-574-5p GAGUGU 4 3.4 GAUAAA 681 hsa-miR-539-5p GAGAAA 4 4.7 CUUUGA 682 hsa-miR-4677-3p CUGUGA 4 3.1 AUUUCU 683 hsa-miR-654-3p AUGUCU 4 2.9 AUUGCG 684 hsa-miR-652-3p AUGGCG 4 7.8 GUUCAA 685 hsa-miR-19a-3p/hsa-miR-19b-3p GUGCAA 4 16.9 GAUGUA 686 hsa-let-7c-5p/hsa-miR-98-5p/hsa-let-7q-5p/hsa-let- GAGGUA 4 1300.2 7f-5p/hsa-miR-202-3p/hsa-let-7b-5p/hsa-let-7e- 5p/hsa-let-7a-5p/hsa-let-7d-5p/hsa-let-7i-5p GAUCAC 687 hsa-miR-3663-3p GAGCAC 4 11.7 CAUUGC 688 hsa-miR-152-3p/hsa-miR-148b-3p/hsa-miR-148a-3p CAGUGC 4 1529.4 GGUGUU 689 hsa-miR-193b-5p GGGGUU 4 2.3 AUUCAC 690 hsa-miR-502-3p/hsa-miR-501-3p AUGCAC 4 5.5 AUUUGG 691 hsa-miR-299-3p AUGUGG 4 4.5 AGUUGU 692 hsa-miR-140-5p AGUGGU 5 4.4 UUGUCA 693 hsa-miR-96-5p/hsa-miR-182-5p UUGGCA 5 25.1 ACUUGC 694 hsa-miR-193b-3p ACUGGC 5 6.9 AAUUCC 695 hsa-miR-365a-3p AAUGCC 5 4.9 CCUUUA 696 hsa-miR-486-5p CCUGUA 5 7.4 CGGUUU 697 hsa-miR-125b-1-3p CGGGUU 5 3.9 UGUUCG 698 hsa-miR-210-3p UGUGCG 5 11.7 GAAUGU 699 hsa-miR-493-3p GAAGGU 5 2.8 AAAUUA 700 hsa-miR-548am-5p AAAGUA 5 3 UGUUCU 701 hsa-miR-218-5p UGUGCU 5 4.4 AAAUUG 702 hsa-miR-20b-5p/hsa-miR-20a-5p/hsa-miR-93- AAAGUG 5 24.7 5p/hsa-miR-17-5p/hsa-miR-106b-5p GGUUGG 703 hsa-miR-541-3p GGUGGG 5 3.8 ACUUUU 704 hsa-miR-452-5p ACUGUU 5 2.4 CCUUGC 705 hsa-miR-221-5p CCUGGC 5 6.3 AAAUCG 706 hsa-miR-518f-3p AAAGCG 5 2 CCUUCU 707 hsa-miR-370-3p CCUGCU 5 6.5 GCAUCA 708 hsa-miR-107/hsa-miR-103a-3p GCAGCA 5 57.4 GGAUUG 709 hsa-miR-122-5p GGAGUG 5 96.6 CCAUCA 710 hsa-miR-338-3p CCAGCA 5 7.3 AAUUUU 711 hsa-miR-409-3p AAUGUU 5 7.8 AAGUCA 712 hsa-miR-124-3p AAGGCA 5 2.7 GAGUUA 713 hsa-let-7d-5p/hsa-let-7q-5p/hsa-let-71-5p/hsa-let-7f- GAGGUA 5 23.9 5p/hsa-let-7e-5p/hsa-let-7a-5p/hsa-let-7b-5p/hsa-let- 7c-5p AGUUCA 714 hsa-miR-130b-3p/hsa-miR-301a-3p/hsa-miR-130a- AGUGCA 5 4 3p/hsa-miR-301b-3p AGUUCU 715 hsa-miR-512-3p AGUGCU 5 2.7 AACUGA 716 hsa-miR-191-5p AACGGA 5 6.8 ACUUCA 717 hsa-miR-509-3-5p ACUGCA 5 38.2 AUUUCA 718 hsa-miR-92a-3p/hsa-miR-92b-3p/hsa-miR-363- AUUGCA 5 41 3p/hsa-miR-25-3 /hsa-miR-32-5 AAGUUG 719 hsa-miR-18a-5p AAGGUG 5 7 AUGUCA 720 hsa-miR-183-5p AUGGCA 5 8.1 GCUUGU 721 hsa-miR-138-5p GCUGGU 5 2.8 CUCUGC 722 hsa-miR-1307-3p CUCGGC 5 2.1 GAGUGG 723 hsa-miR-423-5p GAGGGG 5 3.5 UAAUAC 724 hsa-miR-499a-5p/hsa-miR-208a-3p UAAGAC 5 4.5 CUGUAC 725 hsa-miR-378a-3p CUGGAC 5 8.8 AAAUAA 726 hsa-miR-186-5p AAAGAA 5 3.9 UUUUCA 727 hsa-miR-450b-5p UUUGCA 5 2.3 UUUUCG 728 hsa-miR-450a-5p UUUGCG 5 5.1 GUAUUG 729 hsa-miR-142-3p GUAGUG 5 2.5 ACAUUA 730 hsa-miR-101-3p/hsa-miR-144-3p ACAGUA 5 8.9 AAAUCU 731 hsa-miR-320a AAAGCU 5 6.6 CCAUUG 732 hsa-miR-199b-5p/hsa-miR-199a-5p CCAGUG 5 9.2 UAGUGC 733 hsa-miR-1296-5p UAGGGC 5 2.9 GGAUAG 734 hsa-miR-185-5p GGAGAG 5 2.6 AUGUCU 735 hsa-miR-135a-5p AUGGCU 5 3.1 AGUAUA 736 hsa-miR-411-5p AGUAGA 6 2.5 UCCAUU 737 hsa-miR-145-5p UCCAGU 6 4.2 UCAAUU 738 hsa-miR-26a-5p UCAAGU 6 6.1 ACUGUC 739 hsa-miR-193b-3p ACUGGC 6 2i GGCAUU 740 hsa-miR-34c-5p GGCAGU 6 7.1 GGUAUA 741 hsa-miR-379-5p GGUAGA 6 2.1 CCCUUA 742 hsa-miR-125b-5p CCCUGA 6 3.8 CCUGUC 743 hsa-miR-221-5p CCUGGC 6 3/1 UCUUUA 744 hsa-miR-526b-5p UCUUGA 6 4.4 GGAAUA 745 hsa-miR-7-5p GGAAGA 6 4.4 AGCAUC 746 hsa-miR-16-5p/hsa-miR-15b-5p/hsa-miR-424- AGCAGC 6 4.7 5p/hsa-miR-15a-5p UAAAUC 747 hsa-miR-9-3p UAAAGC 6 4.6 GGGUUG 748 hsa-miR-363-5p GGGUGG AUCUUG 749 hsa-miR-1298-3p AUCUGG 6 11 GAUUUG 750 hsa-miR-509-3p GAUUGG 6 4.4 GCGGUG 751 hsa-miR-744-5p GCGGGG 6 2.1 CAGUUC 752 hsa-miR-148a-3p CAGUGC 6 36.8 AAGUUC 753 hsa-miR-302a-3p AAGUGC 6 9.8 UAGGUC 754 hsa-miR-1296-5p UAGGGC 6 3.3 GAGGUG 755 hsa-miR-423-5p GAGGGG 6 3.7 CUUUUG 756 hsa-miR-9-5p CUUUGG 6 40 GCUGUU 757 hsa-miR-138-5p GCUGGU 6 3.7 AGCUUC 758 hsa-miR-22-3p AGCUGC 6 47 ACUAUA 759 hsa-miR-28-3p ACUAGA 6 3.4 GAUUUU 760 hsa-miR-508-3p GAUUGU 6 8.4 UAUUUC 761 hsa-miR-137 UAUUGC 6 7.2 GGGGUA 762 hsa-miR-5010-5p GGGGGA 6 2 UCUAUA 763 hsa-miR-523-5p UCUAGA 6 2.5 AACGUA 764 hsa-miR-191-5p AACGGA 6 5 CACAUU 765 hsa-miR-128-3p CACAGU 6 2.8 CCAGUU 766 hsa-miR-199a-5p/hsa-miR-199b-5p CCAGUG 7 6.4 CCACUU 767 hsa-miR-181a-2-3p CCACUG 7 5.3 UCACAU 768 hsa-miR-27a-3p/hsa-miR-27b-3p UCACAG 7 5.4 UAUACU 769 hsa-let-7d-3p UAUACG 7 3.3 UUUUUU 770 hsa-miR-129-5p UUUUUG 7 2.2 UGUGCU 771 hsa-miR-210-3p UGUGCG 7 3.6 GAAUUU 772 hsa-miR-219a-2-3p GAAUUG 7 9.2 AAAACU 773 hsa-miR-424-3p AAAACG 7 2.7 AAGGUU 774 hsa-miR-18a-5p AAGGUG 7 5.1 UGAAAU 775 hsa-miR-488-3p UGAAAG 7 2.6 GGAAUU 776 hsa-miR-1-3p GGAAUG 7 3.8 CCAUCU 777 hsa-miR-181a-3p CCAUCG 7 2.4 CAGUAU 778 hsa-miR-199b-3p CAGUAG 7 7.9 ACCCUU 779 hsa-miR-10a-5p ACCCUG 7 3 AGGUAU 780 hsa-miR-196b-5p AGGUAG 7 2.1 GGUUGU 781 hsa-miR-92a-1-5p GGUUGG 7 2.2 AGACGU 782 hsa-miR-483-5p AGACGG 7 2.2 CUGCAU 783 hsa-miR-17-3p CUGCAG 7 2.1 GGGUGU 784 hsa-miR-363-5p GGGUGG 7 2 CUUUGU 785 hsa-miR-9-5p CUUUGG 7 63.3 AAACCU 786 hsa-miR-1537-3p AAACCG 7 2 AAAGUU 787 hsa-miR-106b-5p/hsa-miR-20a-5p/hsa-miR-17- AAAGUG 7 8.4 5p/hsa-miR-93-5p GAGAUU 788 hsa-miR-143-3p GAGAUG 7 33.4 UUUCAU 789 hsa-miR-30a-3p/hsa-miR-30e-3p UUUCAG 7 8.7 GCUCGU 790 hsa-miR-423-3p GCUCGG 7 2.1 AUCUGU 791 hsa-miR-1298-3p AUCUGG 7 3.7 CGACCU 792 hsa-miR-1307-5p CGACCG 7 3.5 GAGGGU 793 hsa-miR-423-5p GAGGGG 7 2.8 GGAGUU 794 hsa-miR-122-5p GGAGUG 7 53.7 UUAUCAU 795 hsa-miR-374a-3p UUAUCAG 8 4.9 GGUGCGU 796 hsa-miR-675-5p GGUGCGG 8 2 GGUUGGU 797 hsa-miR-92a-1-5p GGUUGGG 8 2.1 CGACCGU 798 hsa-miR-1307-5p CGACCGG 8 6.7 AGCAGCU 799 hsa-miR-503-5p AGCAGCG 8 15.3 GGCUCAU 800 hsa-miR-24-3p GGCUCAG 8 4.9 UAUAAAU 801 hsa-miR-340-5p UAUAAAG 8 3.6 ACUGCAU 802 hsa-miR-509-3-5p ACUGCAG 8 6.3 GGCAGUU 803 hsa-miR-34a-5p/hsa-miR-34c-5p GGCAGUG 8 4.2 UCUUGAU 804 hsa-miR-526b-5p UCUUGAG 8 6.3 UGAAAUU 805 hsa-miR-203a-3p UGAAAUG 8 18.8 UGCAUAU 806 hsa-miR-153-3p UGCAUAG 8 3.4 UAAGACU 807 hsa-miR-208a-3p UAAGACG 8 9.8 AAUACUU 808 hsa-miR-200b-3p/hsa-miR-200c-3p AAUACUG 8 15.6 UCUAGAU 809 hsa-miR-518f-5p/hsa-miR-523-5p UCUAGAG 8 6.4 ACUAUAU 810 hsa-miR-625-3p ACUAUAG 8 2.9 GUAACAU 811 hsa-miR-194-5p GUAACAG 8 12.9 UGUACAU 812 hsa-let-7d-3p UGUACAG 8 2.2 AUCUGGU 813 hsa-miR-1298-3p AUCUGGG 8 2.1 ACUCUGU 814 hsa-miR-514a-5p ACUCUGG 8 2.6 AGACGGU 815 hsa-miR-483-5p AGACGGG 8 12 CGUACCU 816 hsa-miR-126-3p CGUACCG 8 13.7 CACAGUU 817 hsa-miR-128-3p CACAGUG 8 12.1 AUACAAU 818 hsa-miR-381-3p AUACAAG 8 7.1 AAAGCUU 819 hsa-miR-320a AAAGCUG 8 4.1 UCUCAAU 820 hsa-miR-513c-5p/hsa-miR-514b-5p UCUCAAG 8 4.3 GCUGGUU 821 hsa-miR-138-5p GCUGGUG 8 3.4 UCCAGAU 822 hsa-miR-520a-5p UCCAGAG 8 4.4 CCCUGAU 823 hsa-miR-125b-5p/hsa-miR-125a-5p CCCUGAG 8 21.3 AACACUU 824 hsa-miR-141-3p AACACUG 8 2.4 UGCCCUU 825 hsa-miR-874-3p UGCCCUG 8 3.5 UCCUAUU 826 hsa-miR-202-5p UCCUAUG 8 22.5 ACCACAU 827 hsa-miR-140-3p ACCACAG 8 3.2 CCCCCAU 828 hsa-miR-361-3p CCCCCAG 8 2 UCACAAU 829 hsa-miR-513b-5p UCACAAG 8 2.5 GUGACUU 830 hsa-miR-134-5p GUGACUG 8 2.9 UGCAUUU 831 hsa-miR-33a-5p UGCAUUG 8 2.1 AGUGCUU 832 hsa-miR-512-3p AGUGCUG 8 2.7 GAGGUAU 833 hsa-let-7a-5p/hsa-let-7c-5p/hsa-let-7b-5p/hsa-let-7d- GAGGUAG 8 36.5 5p/hsa-let-7f-5p/hsa-let-7e-5p/hsa-let-71-5p/hsa-let- 7d-5p UUGUUCU 834 hsa-miR-375 UUGUUCG 8 25.5 AUCAUCU 835 hsa-miR-136-3p AUCAUCG 8 6.3 ACUCCAU 836 hsa-miR-508-5p ACUCCAG 8 10.6 AAGGCACU 837 hsa-miR-124-3p AAGGCACG 9 2.7 UCCCUUUU 838 hsa-miR-204-5p/hsa-miR-211-5p UCCCUUUG 9 2.6 UGUGCGUU 839 hsa-miR-210-3p UGUGCGUG 9 3.8 GAGAACUU 840 hsa-miR-146a-5p/hsa-miR-146b-5p GAGAACUG 9 5.4 GAUUGUAU 841 hsa-miR-508-3p GAUUGUAG 9 4.9 UCACAUUU 842 hsa-miR-23a-3p UCACAUUG 9 6.1 GAAUUGUU 843 hsa-miR-219a-2-3p GAAUUGUG 9 11.4 AACACCAU 844 hsa-miR-21-3p AACACCAG 9 2.7 GGAGUGUU 845 hsa-miR-122-5p GGAGUGUG 9 42.5 UAGAGGAU 846 hsa-miR-877-5p UAGAGGAG 9 2.3 UUUUUGCU 847 hsa-miR-129-5p UUUUUGCG 9 5.8 UCUUGAGU 848 hsa-miR-526b-5p UCUUGAGG 9 4.1 CUUAAACU 849 hsa-miR-302a-5p CUUAAACG 9 13 AUGCCUUU 850 hsa-miR-532-5p AUGCCUUG 9 3.8 UCCAGAGU 851 hsa-miR-520a-5p UCCAGAGG 9 2 CUACAGUU 852 hsa-miR-139-5p CUACAGUG 9 2 ACCCGUAU 853 hsa-miR-99a-5p/hsa-miR-100-5p/hsa-miR-99b-5p ACCCGUAG 9 34.8 AAAGCUGU 854 hsa-miR-320a AAAGCUGG 9 2.6 AAUCUCAU 855 hsa-miR-216a-5p AAUCUCAG 9 5.5 CGGAUCCU 856 hsa-miR-127-3p CGGAUCCG 9 4.3 CGGGUUAU 857 hsa-miR-125b-1-3p CGGGUUAG 9 4.7 AUACAAGU 858 hsa-miR-381-3p AUACAAGG 9 2 UCACAGUU 859 hsa-miR-27a-3p/hsa-miR-27b-3p UCACAGUG 9 5.6 GGGGGAUU 860 hsa-miR-5010-5p GGGGGAUG 9 2.5 UGCCCUAU 861 hsa-miR-3157-3p UGCCCUAG 9 2 CUGCAGUU 862 hsa-miR-17-3p CUGCAGUG 9 2.4 UCUAGAGU 863 hsa-miR-523-5p UCUAGAGG 9 2.8
27. A composition for inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising: the RNA interference-inducing nucleic acid of claim 23.
28. A method of preparing an RNA interference-inducing nucleic acid inhibiting the expression of a non-canonical target gene of microRNA (miRNA) by modifying a partial sequence of specific miRNA in one or more single strands of the double strands of a nucleic acid inducing RNA interference, the method comprising the following steps: constructing an RNA interference-inducing nucleic acid to have a modified base sequence in which at least one guanine base is substituted with an uracil base in a sequence of 6 to 8 consecutive bases, starting from the second base from the 5′ end of specific miRNA.
29. A method of inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising: administrating the composition comprising the RNA interference-inducing nucleic acid of claim 1 into a subject.
30. (canceled)
31. A method of regulating cell cycling, differentiation, dedifferentiation, morphology, migration, division, proliferation or apoptosis, comprising: administrating the composition containing the RNA interference-inducing nucleic acid of claim 1 into a subject.
32. (canceled)
33. A method of inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising: administrating the composition containing the RNA interference-inducing nucleic acid of claim 13 into a subject.
34. (canceled)
35. A method of inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising: administrating the composition containing the RNA interference-inducing nucleic acid of claim 18 into a subject.
36. (canceled)
37. A method of inhibiting the expression of a non-canonical target gene of microRNA (miRNA), comprising: administrating the composition containing the RNA interference-inducing nucleic acid of claim 23 into a subject.
38. (canceled)
Description
DESCRIPTION OF DRAWINGS
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MODES OF THE INVENTION
[0261] Hereinafter, the present invention will be described in further detail with reference to the following examples. The following examples are merely provided to exemplify the present invention, and the present invention are not limited to the following examples.
EXAMPLE 1
Action Mechanism of Interference-Inducing Nucleic Acid Suppressing Non-Canonical Target of miRNA
[0262] Through Ago HITS CLIP assay, the inventors confirmed from an Argonaute protein-binding RNA complex sequence that, when miRNA is base-paired with a target site in Argonaute protein, the miRNA binds with target messenger RNA (mRNA) without making a complete complementary pairing with the miRNA seed, in addition to a canonical seed target having a complete complementary pairing with the miRNA seed. Moreover, as a result of sequencing of RNA having the above-described characteristics, it was confirmed that there are a target base-paired by the induction of a nucleation bulge to the target by the 6.sup.th base of the miRNA (non-canonical nucleation bulge target site of miRNA) and a target site allowing G:A wobble pairing in a miRNA seed region (non-canonical G:A wobble seed target site of miRNA), which naturally occur (
[0263] As illustrated in
[0264] In addition, in the Argonaute-bound RNA complex, a target in which the miR-124 seed base sequence (5′-UAAGGCAC-3′) makes complementary base pairing with a non-canonical nucleation bulge target site (5′-GUGGCCUU-3′) of miRNA to allow G of target mRNA in positions 5 and 6 from the 5′ end of miR-124 to be arranged in a bulge was identified, and based on this, the seed base sequence of miR-124 enabling complementary base pairing with the non-canonical nucleation bulge target site of miRNA has a base sequence (5-UAAGGCCA-3) in which the 6.sup.th base is repeated once more between positions 6 and 7 of the miR-124 seed. The consecutive and perfectly complementary sequence with respect to the above-described sequence was used as miR-124BS (BS: bulge site) siRNA suppressing a non-canonical nucleation bulge target of miR-124.
[0265] When miRNA-bound target mRNA sequence in the Argonaute-bound RNA complex was analyzed, other than a conventional complementary base sequence, it was confirmed that G:A wobble base pairing occurs in the binding of the target mRNA with the seed sequence of miRNA. Therefore, in the case of miR-124, the 4.sup.th G base from the 5′ end of the seed base sequence (5′-UAAGGCAC-3′) makes a G:A wobble pair, thereby recognizing a non-canonical G:A wobble seed target site of miRNA (5′-UGGCAUU-3′). Based on this, siRNA was designed with a consecutively and perfectly complementary sequence with the non-canonical G:A wobble seed target site of miR-124 to invent miR-124-GU, and siRNA was used as siRNA inhibiting the expression of the non-canonical G:A wobble seed target of miR-124.
EXAMPLE 2
Confirmation of Inhibitory Ability of miRNA-BS in Which Non-Canonical Nucleation Bulge Target Site and Consecutively and Perfectly Complementary Base Sequence are Included in Seed Region
[0266] In a canonical seed target site of natural miR-124, the sequence complementary to the 2.sup.nd to 7.sup.th bases from the 5′ end of miR-124 is 5′-GUGCCUU-3′, and the canonical seed target site makes perfectly complementary base pairing with at least 6 consecutive bases with the seed of miR-124 (5′-UAAGGCAC-3′) (
[0267] A non-canonical nucleation bulge target site (Nuc site; nucleation bulge site) of miR-124 found by Ago HITS CLIP assay is 5′-GUGGCCUU-3′, the 6.sup.th base (C) of miR-124 is base-paired with the target by forming a nucleation bulge, and therefore, G of target RNA paired between positions 5 and 6 from the 5′ end of miR-124 is formed in a bulge. Based on this, siRNA was designed using the base sequence having consecutively and perfectly complementary binding with the non-canonical nucleation bulge target site (nucleation bulge site) (5′-UAAGGCCA-3′) as a miRNA seed and named miR-124-BS siRNA (
[0268] First, to confirm how much miR-124 can suppress a non-canonical target, which is a nucleation bulge target site, compared with a conventional canonical target, which is a seed target, a corresponding site for miR-124 was inserted into a luciferase reporter, and various concentrations (0, 0.01, 0.1, 0.5, 2.5 and 15 nM) of miR-124 were also transfected into cells, followed by measuring inhibitory concentration 50 (IC.sub.50) with luciferase activity. As a result, it can be confirmed that the IC.sub.50 of the canonical seed site was approximately 0.5 nM, which was similar to the IC.sub.50 of the non-canonical nucleation bulge site of approximately 0.2 nM (
[0269] The luciferase reporter assay performed herein was conducted by cloning the sequence of the corresponding miR-124 target site into a Renilla luciferase gene (3′ untranslated region (3′UTR)) site of a psi-check2 vector (Promega) twice in a row. The sequence of the non-canonical bulge site uses a naturally-occurring non-canonical nucleation bulge target site in the 3′ UTR site of MINK1. The miR-124, as a human-derived sequence, was prepared as a duplex of a guide strand and a passenger strand, which are chemically synthesized by Bioneer according to a miRBase database, separated by HPLC, according to by the method provided by the manufacturer. The miR-124 and the luciferase reporter vector were co-transfected into approximately 10,000 cervical cancer cells (HeLa: ATCC CCL-2) using a Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's protocol. The HeLa cells were incubated in Dulbecco's modified Eagle's medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 μg/ml streptomycin, and during transfection, the cells were incubated in an antibiotic-free complete medium. Twenty-four hours after transfection, luciferase activity was measured using a dual-luciferase reporter assay system (Promega) according to the manufacturer's protocol, and here, the measurement of Renilla luciferase activity was repeated at least three times using a Glomax Luminometer (Promega), and calculated by normalization by firefly luciferase activity.
[0270] To confirm the functions of miR-124-BS invented to specifically suppress only the non-canonical nucleation bulge target of the miR-124, an experiment was conducted with a luciferase reporter in the same manner as described above. As a result, the miR-124-BS did not inhibit luciferase reporter enzyme activity with respect to the canonical seed target of miR-124 at any concentration (0 to 10 nM), but an effect of inhibiting luciferase reporter enzyme activity with respect to the non-canonical nucleation bulge target reporter of miR-124 was possibly confirmed at a concentration of 0.1 nM or more (
[0271] In this example, the inventors invented miRNA-BS having a consecutive and perfectly complementary seed site sequence with respect to a corresponding site sequence in order to suppress only a non-canonical organic target of miRNA, and when a luciferase reporter assay was performed using miR-124 to verify the effect of miRNA-BS, it was able to be demonstrated that the miR-124-BS does not inhibit the expression of a canonical seed target, and is able to effectively inhibit the expression of a non-canonical bulge target.
EXAMPLE 3
Observation of Apoptosis of Cervical Cancer Cells Induced by Inhibition of Expression of Non-Canonical Nucleation Bulge Target of miR-124
[0272] After confirming that the miR-124-BS suppresses only a non-canonical bulge target among several targets suppressed by miR-124 in the above-described example, through this, it was intended to confirm whether only a specific function of the miR-124 can be exhibited. Since specifically expressed only in nerve cells, the miR-124 is known to mainly play a role related to nerve cells. However, miR-124 expression decreases in gliomas generated by tumors, with regard to this, when the miR-124 expression is artificially induced in tumor cells, it was observed that the apoptosis of tumor cells may occur. Therefore, to confirm whether various and different functions of miR-124 vary according to the type of target, the miR-124-BS was transfected into cervical cancer cells (HeLa), and the apoptosis of the cells was observed through flow cytometry (
[0273] The experiment was performed by transfecting 75 nM of the miR-124 or miR-124-BS duplex into cervical cancer cells (HeLa: ATCC CCL-2) using an RNAiMAX reagent (Invitrogen) according to the manufacturer's protocol, treating the cells with trypsin 72 hours after transfection, detaching the cells from the culture dish, treating the cells using an Annexin V: FITC Apoptosis Detection Kit II (BD Pharmingen) according to the method provided by the manufacturer to detect annexin V for apoptosis and propidium iodide (PI) for necrosis by BD Aria I (BD Biosciences). Here, as a control, a sequence was synthesized to be the same as the sequence of cel-miR-67, which is miRNA of C. elegans.
[0274] As a result of comparing these experimental results for the control, miR-124 expression and miR-124-BS expression, when miR-124 was expressed, it was observed that, in cervical cancer (HeLa) cells, compared with the control, a ratio of progressing cell death by apoptosis (the number of cells stained with annexin V) increases from approximately 30% to approximately 78%, and it was confirmed that the miR-124-BS expression also increased to approximately 73%, which is 2-fold higher than the control (
[0275] It has been reported that the miR-124 is a brain tissue-specific miRNA, and does not have a brain tissue-specific function in other cells not showing tissue cell-specific transcriptome expression (Farh K et al, 2005, Science, 310 (5755) 1817-21). Therefore, to observe a function of the miR-124 that is caused by suppressing a non-canonical bulge target in nerve cells, the miR-124-BS was transfected into a cerebellar neural stem cell line, C17.2, and then cell differentiation was observed in cell morphology (
[0276] In addition, since the miR-124-BS previously induced apoptosis in HeLa cells, whether such a function is also exhibited in nerve cells was observed through flow cytometry for C17.2 cells in the same manner as in Example 2. As a result, in the case of the miR-124 expression, an apoptosis ratio was 22.4%, almost similar to the control which is 21%, and in the case of the miR-124-BS expression, an apoptosis ratio was 33.4%, which is slightly increased (
[0277] Summarizing the results in the above examples, only by inhibition of the expression of a non-canonical nucleation bulge target of miR-124 by the miR-124-BS, the death of cervical cancer cells (Hela) was induced like the miR-124, and therefore, it can be seen that the inhibitory function of the non-canonical bulge target of the miR-124 is the death of cancer cells. In addition, in neural stem cells, as neurite differentiation occurs due to miR-124 expression, little apoptosis and cell cycle arrest occur, and in the miR-124-BS suppressing only a non-canonical bulge target of miR-124, such a function is exhibited in a slightly different form. Accordingly, it can be seen that the nerve cell differentiation function of miR-124 is usually caused by the suppression of a canonical seed target gene which is conventionally known, or in a different form, it was able to be considered that the nerve cell differentiation function of miR-124 may occur by a non-canonical bulge target.
EXAMPLE 4
Confirmation of Neuronal Differentiation in Different Form With Many Branches and Short Neurites (Branched Neurite Outgrowth) by Inhibiting Expression of miR-124 Non-Canonical Nucleation Bulge Target by miR-124-BS
[0278] After confirming the fact that the miR-124-BS induces a different type of neurite differentiation from miR-124 in the previous example, to further examine this, a human neuroblastomas (sh-sy-5y) cell line (CRL-2266) was used for the experiment. Here, in addition to the miR-124-BS, miR-124-BS(4753) designed to have the same miRNA sequence having the same miR-124 seed sequence which is able to recognize and suppress a non-canonical nucleation bulge site of the miR-124, but the other part of which is different was synthesized by the method described in the previous example and used for the experiment. The sequence of the miR-124-BS(4753) used herein is 5′-CAAGGCCAAAGGAAGAGAACAG-3′, and a control was used by being synthesized to be the same as the miRNA of C. elegans, cel-miR-67 (NT; non-targeting). The corresponding miRNA was transfected by the same method as described in the previous example, the cells were cultured for 60 hours, and then the morphological change of the cells was observed using an optical microscope (
[0279] As a result, the miR-124 expression was possible to observe by the morphological change which makes a neurite of the human neuroblastoma (Sh-sy-5y) cell differentiate longer, whereas in the miR-124-BS(4753) having the same seed sequence that can suppress only a non-canonical bulge target of miR-124 as the miR-124-BS, a highly branched neurite outgrowth was observed in one cell (
[0280] Additionally, the neuronal differentiation of the CAD cell line promoted by the miRNA was confirmed by immunostaining for a nerve cell-specific marker, Tuj1 (
[0281] From the result of the above-described example, it can be seen that miR-124 has a function of forming many short neurite branches, which is different from the function of the conventional miR-124 by suppressing only a non-canonical nucleation bulge target. Particularly, there are a different sequence from the miR-124-BS and the same seed sequence, which was observed using the miR-124-BS(4753) capable of suppressing only the non-canonical nucleation bulge target, elucidating the fact that an RNA interference derivative including the seed sequence capable of recognizing the non-canonical nucleation bulge target exhibits the same effect as the miR-124-BS. In addition, it can be shown that complex neurite branches generated by the miR-124-BS can be applied as a method of promoting various types of neuronal differentiation similar to human brain cells showing a complicated neural network.
EXAMPLE 5
Confirmation of Morphological and Molecular Biological Features of Branched Neurite Outgrowth Induced Thereby After miR-124-BS is Expressed Using siRNA and shRNA Vector
[0282] To additionally confirm the specific branched neurite outgrowth phenomenon induced by the miR-124-BS expression in mouse neuroblastomas N2a (CCL-131) cells, as in Example 4, the miR-124 and the miR-124-BS(4753) were transfected into the cells, and then the morphological change of the cells was observed while the cells were incubated for 72 hours (
[0283] As a result, in the experiment group into which the miR-124 is delivered, nerve cells having a long neurite similar to that in catecholamine-containing nerve cells, CAD, were able to be observed, and in cells to which miR-124-BS(4753) was transfected, the morphological feature of a cell having a short neurite branched in various directions was possible to confirm (
[0284] In all of the examples described above, the miRNA expression was performed by transfecting a synthesized duplex into cells, or also by transfecting a vector for expressing the form of shRNA, which is a hairpin-shaped RNA capable of generating the miRNA. Therefore, a vector for expressing miR-124, miR-124-BS(4753) or miR-124(3714) in the form of shRNA was formed using a pCAG-miR30 plasmid (Addgene #14758), which is a vector for expressing pre-miRNA. The pCAG-miR30 vector used herein used a CAG promoter to strongly express miRNA, was designed to express a backbone of miRNA-30, thereby having an advantage of maximizing miRNA expression (Paddison P et al, Nature methods, 2004, 1(2): 163-7).
[0285] The pCAG vector formed to express each of the miR-124, miR-124-BS(4753) and miR-124(3714) was transfected into N2a cells, using a pCAG vector not expressing anything, and then cell morphology was observed (
[0286] From the results in the example, it can be seen that, even when a vector expressing the miRNA transfection in the form of shRNA, if only having the same seed sequence of the miR-124 (miR-124(3714)), like the miR-124, generally long neurite differentiation occurs by the inhibition of the expression of a canonical target gene. In addition, when the non-canonical bulge target of miR-124 is also expressed in the same manner as the miR-124-BS through the seed region in the form of shRNA (miR-124-BS(4753)), branched neurite outgrowth may occur. It was possible to confirm that both of the canonical target suppression and non-canonical bulge target suppression by the miR-124 show molecular gene marker expression in the form of neurite differentiation of nerve cells, and both show differentiation into nerve cells.
EXAMPLE 6
Confirmation of Induction of Neurite Differentiation of Mouse Neuroblastoma (N2a) Cells by miR-124-BS Through Regulation of MAPRE1 Expression
[0287] Additionally, the miR-124, miR-124-B S(4753) and miR-124(3714)-induced neurite differentiation in neuroblastoma (N2a) cells was observed in terms of cell morphology, which was quantitatively analyzed (
[0288] As a result, with respect to 100 cells, the number of neurite branches expressed per cell was increased in the miR-124, miR-124-BS(4753) and miR-124(3714) experimental groups approximately 3- to 5-fold compared to the control (
[0289] To investigate whether the specific neuronal differentiation phenomenon caused by the miR-124-BS is induced by a non-canonical bulge site of any gene, the non-canonical bulge target sequence of miR-124 was searched for in the sequence of the 3′ UTR region of the gene related to neuronal differentiation. As a result, a microtubule-associated protein RP/EB family member 1 (MAPRE1: Elena Tortosa et al. 2013 EMBO32,1293-1306) gene reported to regulate the differentiation of nerve cells was found. MAPRE1 may be a protein attached to the end of a microtubule constituting a neurite in the generation of the neurite, and may determine the shape of a neurite through the regulation of the formation of the corresponding microtubule. Accordingly, to confirm that the MAPRE1 gene is recognized as the non-canonical bulge target of miR-124 and inhibits the expression thereof, after miR-124-BS was transfected into N2a cells, the mRNA level of MAPRE1 was measured by qPCR assay according to the method performed in Example 5 (
[0290] MAPRE1 is the non-canonical bulge target of the miR-124, and if it is a very important target, the type of neuronal differentiation caused by the suppression of the non-canonical bulge target of the miR-124-BS is further promoted by the reduction in MAPRE-1 expression. Accordingly, to confirm this, two types of siRNA capable of suppressing MAPRE1 were prepared and transfected into N2a cells with miR-124, followed by performing an experiment of confirming a change in cell morphology (
[0291] Therefore, it was confirmed that the short, branched neurite outgrowth induced by the miR-124-BS is a biological function that can be shown by at least partially inhibiting the expression of a MAPRE1 gene by the non-canonical bulge target site of miR-124.
EXAMPLE 7
Analysis of Tendency of Suppressing Non-Canonical Bulge Target Gene of miR-124 When miR-124-BS is Transfected Into Cells at Transcriptome Level
[0292] It was confirmed that a gene suppressed by miR-124-BS in mouse neuroblastoma (N2a) cells eventually induces the nerve cell differentiation which produces many neurite branches of neuroblastoma (
[0293] After miR-124 and miR-124-BS were transfected into N2a cells, RNA-Seq assay was performed on the miR-124-BS invented by the inventors to confirm only the non-canonical bulge target gene of miR-124 at the transcriptome level expressing all genes. Here, as a control, a duplex having the same base sequence as miR-124 and an abasic substitution (pi) in position 6 from the 5′ end (miR-124-6pi) was used, and it was reported that such substitution does not make pairing with target mRNA at all since there is no base in position 6 from the 5′ end of miRNA (Lee H S et. Al. 2015, Nat Commun. 6:10154).
[0294] The RNA-Seq assay was performed by delivering each of the miR-124, the miR-124-BS and the miR-124-6pi duplex into N2a cells at 75 nM using a modification of the sequence of cel-miR-67, which is miRNA of C. elegans as an experimental control, in which the position 6 from the 5′ end is substituted with an abasic spacer (NT-6pi), extracting total RNA with an RNAeasy (Qiagen) 24 hours after incubation, constructing a library at Otogenetics, and subjecting it to next-generation sequencing. Afterward, a FASTAQ file, which is the sequence data obtained from the assay, was mapped to a mouse genomic sequence (mm9) with the TopHat2 program, expression values (FPKM) were obtained with Cufflink and Cuffdiff programs and normalized to a result in mouse neuroblastomas (N2a) cells into which a control NT-6pi was transfected to perform analysis with a fold change, (log2 ratio).
[0295] To analyze whether the mRNA level of a gene having a target site of miRNA is reduced by the expression of the corresponding miRNA from the RNA-Seq result, genes with a phastCons score of 0.9 or more were screened to select those in which a canonical seed site (7mer paired with bases between positions 2 to 8 from the 5′ end) of the miR-124 has 3′ UTR and the sequence of the corresponding site is conserved in various species, and these profiling results were compared and analyzed with a cumulative fraction in the order of inhibition depending on the expression of the corresponding miRNA. In addition, the non-canonical bulge site (nuc, 7mer) of miR-124 was also identified in the same manner as described above, and the suppressive fraction of the corresponding gene was analyzed with a cumulative fraction in the same manner as described above. Here, since a gene having the non-canonical bulge site of the miR-124 also has a canonical seed site, it is difficult to determine the suppressive effect thereof, in contrast to when there is no canonical seed sequence of miR-124 in total mRNA (nuc only), the case where there is only a canonical seed site and no non-canonical bulge site (seed only) was also analyzed.
[0296] In the RNA-Seq sequencing for the miR-124-expressed experimental group, when a fold change according to the miR-124 expression was analyzed according to a cumulative fraction, a phenomenon in which a gene (miR-124 seed) having the canonical seed site conserved in the miR-124 in the 3′ UTR is highly suppressed compared with the distribution of total mRNA was confirmed (
[0297] When the miR-124-BS developed by the inventors was expressed, in RNA-Seq sequencing, when a fold change according to the miR-124-BS expression was analyzed according to a cumulative fraction, it was confirmed that a weak inhibitory effect was shown in a gene (miR-124 seed) having the canonical seed site conserved in the miR-124 in the 3′ UTR, but there is no change in a gene (miR-124 seed only) having the corresponding canonical seed site without a non-canonical bulge site of the miR-124 (
[0298] From the result of the example, it was able to be seen that miRNA very strongly and effectively suppresses a conventional canonical seed target gene, but very weakly suppresses the non-canonical bulge target at the transcriptome level, and it was possible to confirm that miRNA-BS inhibits the expression of the non-canonical bulge target of miRNA, but does not suppress the conventional canonical seed sequence at the transcriptome level.
EXAMPLE 8
Confirmation of Cell Cycle Arrest Induced by miR-122-BS in Human Liver Cancer Cell Line (HepG2) Through Flow Cytometry
[0299] Based on the result of observing the induction of nerve cell differentiation of miR-124 by the non-canonical nucleation bulge target of miR-124, an experiment for the biological function of the non-canonical nucleation bulge target of another miRNA was performed. MiR-122 has 5′-UGG AGU GU-3′ as a seed base sequence, and miR-122-BS, which is the base sequence of siRNA base-paired with the non-canonical nucleation bulge target site thereof, may have one more U in position 6, and in this case, thus have 19 bases represented by 5′-UGG AGU U GUG ACA AUG GUG-3′ at the 5′ end thereof and deoxy thymine nucleotides (dts) as bases in positions 20 and 21. Particularly, in a duplex structure, the corresponding dt parts may consist of a guide strand and a passenger strand to form a two-nucleotide overhang at the 3′ end. Here, the passenger strand made perfectly complementary base pairing with the guide strand, and included two 2′OMes at both of the positions 1 and 2 from the 5′ end and two dts at the end of the base sequence (
[0300] The miR-122-BS prepared as described above (
[0301] As a result, compared with the cell cycle analysis for the NT-6pi-transfected control, in the miR-122-BS experimental group, the G0/G1-phase cells increased approximately 2-fold from 10.7% to 24.3%, confirming that the cell cycle arrest in the liver cancer (HepG2) cells increased (
[0302] Accordingly, it was possible to observe that the miR-122-BS shows a biological function of inducing cell cycle arrest in a human liver cancer cell line through regulation of the expression of the non-canonical nucleation bulge target of the miR-122, which is a completely different function from the biological function of the miR-122.
EXAMPLE 9
Confirmation of Muscle Fibrosis Function in Skeletal Muscle Cells (C2C12), Induced by miR-1-BS, and Apoptosis Mediated by miR-155-BS
[0303] Since a novel biological function different from the function shown by the miRNA-BS-mediated inhibition of the expression of the canonical seed target of miRNA has been observed, to observe how such a function is exhibited in muscle cells, miRNA-BS was applied to miR-1 and miR-155, which are expressed in muscle cells and play a critical role.
[0304] First, since the miR-1 is a muscle tissue-specific miRNA, and has been reported to function in muscle cell differentiation (Chen J et.al, Nature genetics, 2006, 38(2): 228-233), miR-1-BS was expressed in the skeletal muscle cell line, C2C12, and then a cell morphological change (
[0305] As a result, when miR-1-BS was expressed under the growth media (GM) condition in the mouse skeletal muscle cells (C2C12), compared with the NT (control) or miR-1 duplex-transfected cells, a larger number of differentiated muscle cells was possible to confirm by MF20 staining. Under the deprivation media (DM) condition that reduces serum during incubation, the progression of the differentiation into a muscle fiber was observed, and it was possible to observe a fiber in a thicker form compared with the skeletal muscle cells expressing NT as a control (
[0306] The miR-155 is miRNA inhibiting muscle cell differentiation (Seok H et. Al, JBC, 286(41):35339-46, 2011), the effect of miR-155-BS was observed through the differentiation of skeletal muscle cells (C2C12) in the same manner as performed for the miR-1 above (
[0307] In this example, it was possible to confirm that the miR-1-BS promotes the differentiation of skeletal muscle cells and induces the differentiation of a thick muscle fiber, the miR-155-BS induces the death of skeletal muscle cells, and these functions are different from the function of the conventional miR-1 for contracting muscle cells and the function of the miR-155 for inhibiting muscle cell differentiation.
EXAMPLE 10
Discovery of Non-Canonical Target Site Allowing Wobble Base Pairing at Seed Position of miRNA Through Ago HITS CLIP Assay
[0308] As a method of analyzing a miRNA target at the transcriptome level, an experimental method called Ago HITS CLIP has been developed and is widely used (Nature, 2009, 460 (7254): 479-86). The Ago HITS CLIP assay is a method of inducing covalent bonding between RNA and Argonaute protein in cells by UV irradiation of cells or a tissue sample, isolating the generated RNA-Argonaute complex through immunoprecipitation using an antibody specifically recognizing Argonaute, and analyzing the isolated RNA through next-generation sequencing. The base sequence data obtained thereby may not only identify target mRNA of miRNA through bioinformatics analysis, but also precisely analyze its binding site and sequence (Nature, 2009, 460 (7254): 479-86). Ago HITS-CLIP was first applied to mouse cerebral cortex tissue to perform mapping for the miRNA target in the brain tissue. Since then, this method has been applied to various tissues. Through numerous Ago HITS-CLIP assays, miRNA binding sites in various tissues were identified, and it was found that there are many non-canonical binding sites which make similar miRNA seed binding particularly in the above-mentioned target sequence, but different binding in the sequence of target mRNA (Nat Struct Mol Biol. 2012 Feb. 12; 19(3):321-7).
[0309] It was observed that some of the known non-canonical binding rules are present as wobbles through G:U base pairing, other than the conventional known base pairings. The wobble base pairing is found in specific RNA, different from the conventional base pairing, and it was shown that the well-known G:U wobble pair may also be paired to a non-canonical target of miRNA. However, compared with this, a G:A wobble pair which has weak binding and is not often found may form base pairing in specific RNA, but other than this, has not been investigated at all. However, in the case of the seed sequence of miRNA, since the free energy of base pairing is structurally stabilized by the Argonaute protein, generally weak G:A base pairing may be relatively significant, and based on fact described above, the inventors conducted analysis as follows.
[0310] First, to confirm whether wobble base pairing including a G:A pair is present in the miRNA target of human brain tissue, Ago HITS-CLIP data from the gray matter of the brain tissue after death was analyzed (Boudreau R L et al, Neuron, Vol. 81 (2), 2014, 294-305) (
[0311] As a result of the analysis, all of G:A wobble pairs pair-allowed target sites (median: 1119.5 sites, error range: +/−526.48) and G:U wobble pair-allowed target sites (median: 995 sites, error range: +/−523.22) showed higher distribution than the control (median 891 sites, error range: +/−410.71), indicating that, particularly, there are approximately 1.1 times more miRNA target sites having a G:A wobble pair than those having a G:U wobble pair (
[0312] In this example, as it was found that the recognition of the non-canonical target of miRNA allows G:A wobble pairing to the seed sequence, it is considered that many miRNAs may bind to the mRNA of a target gene, thereby inhibiting the expression of the gene.
EXAMPLE 11
Confirmation of Development of mIRNA-GU Recognizing G:A Seed Site, Which is Non-Canonical Target of miRNA, and Effect of Increasing Nerve Cell Death of miR-124-G5U When miR-124 is Applied
[0313] In Example 10, it was found that the non-canonical target recognition of miRNA may allow a G:A wobble pair in the seeding sequence, which is referred to as a non-canonical G:A seed site, and in contrast, miRNA-GU, which is the sequence of a new RNA interference-inducing material capable of complementarily pairing was invented. When applying such sequencing technology to miR-124, the miR-124 is the base sequence in which a seed sequence is 5′-AAGGCAC-3′ and a non-canonical G:A wobble seed target is capable of having G:A wobble base pairing at the 4.sup.th and 5.sup.th G bases, and therefore, when such a G:A wobble pair is changed into the miRNA-GU sequence, which is a format for changing the wobble pair into a conventional complementary base pair, the sequence may be changed into miR-124-G4U (5′-AAUGCAC-3′) or miR-124-G5U (5′-AAGUCAC-3′). Since the miR-124-G4U and miR-124-G5U changed as described above may be complementarily paired with a non-canonical target site weakly paired in a G:A wobble pair and recognized by the conventional miR-124, the function of the non-canonical G:A seed target in the miR-124 may be exhibited.
[0314] Accordingly, an experiment was performed by applying a non-canonical G:A wobble seed target to the miR-124 to examine the biological function mediated by the non-canonical G:A wobble seed target of the miRNA. To this end, NT (referred to as N2a of
[0315] Based on this, it was possible to confirm that the miR-124-GU-mediated suppression of the non-canonical G:A wobble seed target eliminates the neuronal differentiation ability induced by miR-124, and exhibits an apoptosis regulating function, which is another biological function, instead of the neuronal differentiation ability.
EXAMPLE 12
Promotion of Cell Division of Neuroblasts Induced by miR-124-G4U
[0316] In the previous example, since the miR-124-G4U lost neural differentiation ability and did not have a specific function in apoptosis, cell division was examined. That is, each of the control NT, the miR-124, the miR-124-G4U and the miR-124-G5U was transfected into neuroblastoma (N2a) cells in the same manner as described in the previous example, and flow cytometry for the cell division and proliferation phenomenon was performed (
[0317] Additionally, to investigate an effect of miR-124-G4U on a cell cycle, flow cytometry was performed using a human neuroblastomas cell line (sh-sy-5y) (
[0318] According to the above-described example, it was able to be seen that the miR-124-G4U of the RNA interference-inducing modified sequences suppressing the non-canonical G:A wobble seed target of miR-124 has a function of increasing cell division and proliferation of neuroblasts.
EXAMPLE 13
Confirmation that miR-1 can Inhibit the Expression of a Non-Canonical G:A Wobble Seed Target Gene in a Cardiomyocyte Cell Line Using a Fluorescent Protein Reporter
[0319] In the above-described example, the miRNA was able to non-canonically bind with target mRNA by allowing a G:A wobble pair in a seed sequence, and it was found that the resulting complex was able to have a novel function, and then a reporter assay for confirming whether such binding can actually induce the suppression of a target gene was conducted at the cellular level. Here, this assay was conducted on miR-1 known to be highly expressed in muscle-like cells such as cardiomyocytes h9c2, and particularly, on a G:A wobble base pair, focusing on G present in position 7 from the 5′ end of miR-1 (
[0320] A reporter was formed by inserting a non-canonical target site (7G:A-site) which is complementary to the bases in positions 2 to 8 from the 5′ end of miR-1 and has a G:A wobble at the 7.sup.th base (G) into the fluorescent protein expression reporter vector constructed as described above, and then whether the reporter is suppressed by miR-1 was confirmed through an experiment. Here, in this experiment, 500 ng of the GFP-7G:A site reporter vector and 25 μM of miR-1 were co-transfected into a cardiomyocyte cell line (H9c2) using a Lipofectamine 2000 (Invitrogen) reagent according to the manufacturer's protocol, and 24 hours after transfection, each fluorescent signal was measured through flow cytometry using Attune NxT (Life Technology). As a result, it was confirmed that the 7G:A site of the miR-1 was very effectively suppressed by miR-1 expression (
[0321] In the control, it was observed that the GFP-7G:A site reporter vector was inhibited to some extent even in the h9c2 cells without miR-1 transfection. It may be predicted that the inhibition is mediated by miR-1 previously expressed in the h9c2 cells through G:A wobble base pairing with the reporter target mRNA. To confirm this, a miRIDIAN microRNA hairpin inhibitor (miR-1 inhibitor,
[0322] This was confirmed by additionally transfecting each of a control (GFP-no site) in which a miRNA target site was not added to a fluorescent protein expression reporter and a miR-1 7G:A site-inserted reporter, that is, a GFP-7G:A site into the H9c2 cells and comparing their activities, and the result was quantitatively compared with the case of co-transfection with miR-1 as a positive control (
[0323] To confirm that the result observed in the above example was not influenced by the intensities of different promoters used in the corresponding fluorescent protein reporter and the difference in fluorescent activity between two different fluorescent proteins, a fluorescent protein reporter, that is, p.UTA.3.0 (Lemus-Diaz N et al, Scientific Reports, (7), 2017), in which two fluorescent proteins were interchanged was purchased from Addgene (plasmid #82447). Here, a reporter assay was conducted after constructing a reporter vector (RFP-7G:A site) by repeatedly inserting the miR-1-7G:A sequence five times into the 3′ UTR of the RFP regulated by a SV40 promoter in a p.UTA.3.0 vector, and the RFP expression level was used by modifying a GFP value normalized to a degree of transfection into cells (
[0324] From the above, it was confirmed that the non-canonical target of miRNA found by Ago HITS-CLIP assay cannot only actually bind to the seed sequence of miRNA through G:A wobble pairing, but also inhibits the expression of the corresponding target gene. According to this, it is considered that the non-canonical target of miRNA is able to easily bind to the target site of miRNA through G:A wobble pairing, and if this can be similarly induced by miRNA-GU, only the biological function of miRNA mediated by the non-canonical target suppression will be used.
EXAMPLE 14
Confirmation of Function of Inducing the Hypertrophy of Cardiomyocytes Through Regulation of Non-Canonical G:A Wobble Seed Target Gene at G2, G3 and G7 of miR-1
[0325] To confirm the possibility of regulating a biological function through the above-described non-canonical gene suppression by the G:A wobble base pair acting as a non-canonical target of miRNA by using miRNA-GU, an experiment was conducted with the miR-1 of a cardiomyocyte, which has been described in the previous example.
[0326] To investigate whether a non-canonical G:A wobble target has a specific biological function, miR-1 should recognize only a non-canonical G:A wobble pair target site, rather than a canonical target, and thus miRNA-GU prepared by substituting the corresponding G with U in the seed region of the miR-1 to be paired with A, not C, was used for the experiment. In this experiment, for physiological conditions more similar to the heart, primary rat neonatal cardiomyocytes derived from rat cardiac tissue were used. Here, for cardiomyocyte culture, cardiomyocytes were isolated from cardiac tissue of a neonatal rat (1 day after birth) through an enzyme reaction (Neomyt kit, Cellutron), and the cardiomyocytes were cultured in a 4:1 mixture of Dulbecco's modified Eagle's medium (Invitrogen) supplemented with 10% FBS (fetal bovine serum), 5% HS (horse serum), 100 U/ml penicillin and 100 μg/ml streptomycin and M199 (WellGene). By adding brdU to the prepared media, the cells were cultured differently from other cells with cell cycles in cardiac tissue.
[0327] It was first confirmed whether myocardial hypertrophy is induced in the primarily cultured cardiomyocyte obtained as described above (
[0328] MiR-1 contains three Gs in the seed region, and therefore, to recognize only a non-canonical G:A wobble pair target site, not a canonical target, miR-1 in which G is substituted with U was designed and applied to the 2.sup.nd position (miR-1-G2U), the 3.sup.rd position (miR-1-G3U) and the 7.sup.th position (miR-1-G7U) from the 5′ end depending on where G is located. Here, the substituted base sequences of the miR-1 used in
[0329] As a result, it was possible to observe that when miR-1-G2U, miR-1-G3U or miR-1-G7U was transfected into the primary cardiomyocyte culture, the size of the cardiomyocytes was increased similar to the myocardial hypertrophy-induced cells (
[0330] From the above, it was finally seen that the non-canonical target suppressed by the G:A wobble pair in the miRNA seed region can exhibit a biologically different function from the conventional canonical target recognition. This finding indicates that the function caused by a canonical target of miRNA is different from the function caused by a non-canonical target thereof, and according to this, if only the G:A wobble target of the miRNA can be suppressed, only a biological function occurring due to the G:A wobble target will be selectively controlled.
EXAMPLE 15
Observation of Induction of Myocardial Hypertrophy Through miR-133-G4U Expression in Cardiomyocyte Cell Line (H9c2)
[0331] In terms of the non-canonical target phenomenon recognized by G:A wobble base pairing acting on the seed region of miRNA, to additionally confirm its biological function in cardiomyocytes, other than the miR-1 described in the previous example, miRNA-GU was applied to miR-133 known to function in cardiomyocytes. The miR-133 is known to have a function of regulating myocyte development and disease pathology and inhibiting myocardial hypertrophy (Nat Med. 2007, 13(5): 613-618; Ikeda S et al, Mol cell boil, 2009, vo129, 2193-2204). Therefore, the inventors transfected a 50 nM duplex prepared with miR-133-G4U (5′-UUUUGUCCCCUUCAACCAGCUG-3′), which is an interference-inducing nucleic acid specifically inhibiting target recognition through the G4 from the 5′ end of a non-canonical GA wobble target site of the miR-133, synthesized by Bionia as described above into a cardiomyocyte cell line (h9c2), and observed a change in cell morphology (
[0332] Accordingly, it was possible to predict that the decrease in expression of the non-canonical G:A wobble seed target of miR-133, occurring by miR-133-G4U induces myocardial hypertrophy, which is a novel function different from the canonical seed target inhibitory function of miR-133 for inhibiting myocardial hypertrophy.
EXAMPLE 16
Confirmation of Effect of Inhibiting Expression of Non-Canonical GA Wobble Seed Site Gene by miR-122 in Liver Cancer Cells Using Luciferase Reporter
[0333] In Example 10 described above, from the finding that miRNA non-canonically binds with a G:A wobble pair seed site, miRNA-GU specifically recognizing only the non-canonical target binding was invented, and it was observed in Examples 11, 12, 13 or 14 that, when this was applied to miR-124, miR-1 or miR-133, an effect different from the conventional function was exhibited. In addition, to confirm whether the expression of a target gene having a non-canonical G:A wobble pair seed site in miRNA is able to be actually inhibited, the function in a myocardial tissue cell line was confirmed for miR-1 in Example 13. In addition, it was attempted to confirm whether the expression of a non-canonical G:A wobble seed target gene can be inhibited in miR-122 specifically expressed in liver tissue and liver cancer cells, using a luciferase reporter system (
[0334] The miR-122 specifically expressed in liver cancer or liver tissue and thus functioning has Gs capable of making a G:A wobble pair in positions 2, 3, 5 and 7 from the 5′ end of the seed sequence (the 1.sup.st to 8.sup.th bases from the 5′end: 5′-UGG AGU GU-3′). Therefore, first, in order to measure the inhibitory efficiency of the corresponding target site capable of being non-canonically recognized through G:A wobble pairing at G2, an inhibitory concentration 50 (IC50) was measured by the same method as described in Example 2 and compared with a canonical seed site (
[0335] To detect the inhibition of the expression of a non-canonical G:A wobble seed target enabling G:A base pairing with G2 of miR-122, a luciferase reporter vector for an experiment was produced by introducing five sequentially arranged copies of a non-canonical G:A wobble seed site (2G:A) sequence (the 1.sup.st to 9.sup.th bases from the 5′ end: 5′-CAC ACU CAA-3′) for the G2 of miRNA-122 into the 3′ untranslated region (3′ UTR) of a Renilla luciferase gene in a psi-check2 (Promega) vector In addition, a luciferase reporter vector for a canonical seed site (the 1.sup.st to 9.sup.th bases from the 5′ end: 5′-CAC ACU CCA-3′) was simultaneously produced in the same manner as described above.
[0336] As a result of measuring an effect of inhibiting the expression of a target gene having a canonical seed site and a non-canonical G:A wobble seed site (2G:A site) at G2 from the 5′ end at various concentrations of miR-122 by a change in activity of a corresponding luciferase reporter using the luciferase reporter vector produced as described above, and examining an IC50 value, the IC50 of the miR-122 was measured to be approximately 0.5 nM for a canonical seed target site, and to be approximately 7 nM for a non-canonical G:A wobble seed site (2G:A site), confirming that the miR-122 inhibits the expression of the gene having the non-canonical G:A wobble seed site (2G:A site), and showing that the efficiency is lower compared to the canonical seed site (
[0337] This experiment was conducted by synthesizing a duplex of miR-122 in the same manner as used in this example, co-transfecting it with a corresponding psi-check2 vector (50 ng) at various concentrations (0, 1, 5, 10 and 25 nM) into approximately 10,000 liver cancer cells (Huh7, KCLB: 60104) according to the manufacturer's protocol using a Lipofectamine 2000 reagent (Invitrogen), and incubating the cells in a 96-well plate, and then luciferase activity was measured in the same manner as used in Example 2.
[0338] After confirming that miR-122 inhibits the expression of the non-canonical G:A wobble seed site gene, to confirm whether it is equally regulated by miR-122 present in cells, using a Huh7 cell line, which are liver cancer cells containing miR-122, a luciferase reporter assay was performed on a non-canonical G:A wobble seed site (2G:A) caused by G2 and the G3 from the 5′ end (
[0339] As a result, when a luc-PM site vector was transfected into Huh7 cells known to have miR-122, it was observed that the activity was reduced by approximately 50% compared with the control into which only a luciferase vector was transfected, confirming that there is miR-122 in Huh7 cells, and simultaneously, the miR-122 suppresses the activity of a non-canonical G:A wobble seed target reporter (Luc-3G:A) at G3, and non-canonical G:A wobble seed target reporters (Luc-2G:A, 3G:A) at G2 and G3. In addition, to see whether the suppressive effect is induced by the miR-122, an miR-122 expression inhibitor (hsa-miR-122-5p inhibitor, IH-300591-06-0010) was purchased from Damacon and treated at 50 nM, confirming that all inhibitory phenomena disappeared.
[0340] According to the example, it was confirmed that the miR-122 is weaker than a canonical seed site, and can inhibit the expression of the non-canonical G:A wobble seed target caused by G2 from the 5′ end, and miR-122 naturally present in liver cancer cells (Huh7) inhibits the expression of the non-canonical 3G:A wobble seed target gene and the non-canonical 2,3G:A wobble seed target gene based on the 5′ end. Particularly, the inhibitory effect shown in the liver cancer cells (Huh7) disappears due to the treatment of the miR-122 expression inhibitor, which is regulated by miR-122 (
EXAMPLE 17
Confirmation of Phenomenon of Inhibiting Cell Migration in Liver Cancer Cell Line Through Inhibition of Expression of Specific Non-Canonical G:A Wobble Seed Target Gene of miR-122
[0341] To examine the function of a non-canonical G:A wobble seed target of miR-122 in liver cancer cells, miRNA-GU complementarily recognizing and suppressing the corresponding non-canonical G:A wobble seed target site was applied to be transfected into liver cancer cells (hepG2), and then a wound healing assay was performed to examine how the cell migration ability among the properties of the liver cancer cells, which is important for cancer metastasis, changes.
[0342] The wound healing assay for the liver cell line (hepG2) was performed by first synthesizing miRNA-GU complementarily recognizing the non-canonical 2G:A wobble seed target, the non-canonical 3G:A wobble seed target, or the non-canonical 2,3G:A wobble seed target based on the 5′ end of miR-122 with a corresponding sequence, transfecting the resulting sequence into the hepG2 cells in the same manner as described in the above-described example, making a scratch in a cell layer using a 1,000-μl tip 24 hours after culture, and incubating the cells until 48 hours and observing cell migration in an experimental group, and comparing with cells transfected with a control NT-bpi. The sequences of interference-inducing nucleic acids used in the assay are as follows: (miR-122: 5′-UG GAG UGU GAC AAU GGU GUU UG-3′; miR-122-G2U: 5′-UU GAG UGU GAC AAU GGU GUU UG-3′; miR-122-G3U: 5′-UG UAG UGU GAC AAU GGU GUU UG-3′; miR-122-G2,3U: 5′-UU UAG UGU GAC AAU GGU GUU UG-3′).
[0343] As a result, in the case of the control NT-6pi and the miR-122-transfected case, in 48-hour cell culture, it was possible to observe the migration of cells almost completely filling the scratch. However, in experimental groups transfected with miR-122-G2U and miR-122-G2,3U siRNA, it was seen that the cell migration is inhibited, and such an inhibitory phenomenon is most strongly shown in the miR-122-G2,3U siRNA-transfected experimental group (
[0344] Based on this, it can be seen that the miR-122-mediated suppression of a non-canonical G:A wobble seed target inhibits the migration of liver cancer cells, indicating that G:A wobble at the 2.sup.nd base preferentially acts for inhibition, and when the G:A wobble at the 3.sup.rd base is added, the intensification of the cell migration inhibitory function is induced.
EXAMPLE 18
Confirmation of Cell Cycle Arrest by Regulation of miR-122-G2,3U-Mediated Non-Canonical G:A Wobble Seed Target
[0345] Cell migration is closely related to cell division, and widely observed, particularly, in cancer cells (Cancer research, 2004, 64(22):8420-8427). Accordingly, to confirm whether the miR-122-G2,3U-induced inhibition of liver cancer cell migration is related to the regulation of a cell cycle, flow cytometry was performed in the same manner as in Example 12. Here, as a result of measuring the cell cycle of each type of cells after NT-6pi, miR-122, miR-122-G2U, miR-122-3U and miR-122-G2,3U were delivered into the liver cancer cell line (HepG2), in the miR-122-G2,3U-transfected experimental group, it was confirmed that the ratio of the cells in the G0/G1 phase is increased by 52% to 68% on average compared with the control (NP-6pi), and the G2/M-phase cell distribution is significantly low (
[0346] Accordingly, it was confirmed that the cancer cell function regulated by miR-122-G2,3U reduces cell division (G2/M), and induces cell cycle arrest (G0/G1), and the result of the example may be interpreted as miR-122 recognizes non-canonical GA wobble seed sites through G:A wobble pairing at both of G2 and G3 based on the 5′ end, and exhibits the function of preventing the progression of a liver cancer cell cycle.
EXAMPLE 19
Observation of Inducing Cell Cycle Arrest by Regulation of miR-122-G2,3U, miR-122-G2,7U-Induced Non-Canonical G:A Wobble Seed Targets
[0347] The miR-122 function at a non-canonical G:A wobble seed target identified that miR-122-G2,3U has a function of inhibiting liver cell migration in Example 17, and particularly, it was observed that the inhibition of cell migration induced by the expression of the miR-122-G2,3U is maximized when a regulatory effect of G3 is added to the biological function of the 2.sup.nd base for regulating a non-canonical G:A wobble seed target (
[0348] Here, for the experiment, a liver cancer cell line (HepG2) was used in the same manner as described above, and a corresponding miRNA-GU duplex was prepared in the same manner as in Example 18 above and transfected into the cells at 50 nM. However, in this cell cycle observation, after 24-hour culture, the cells were treated with 100 mg/mL of nocodazole for 16 hours, HepG2 cells at the G2/M phase (division preparation phase/division phase) was synchronized, and the amount of cells in a cell cycle arrest state at G0/G1 was measured using a Muse Cell Analyzer (Millipore) according to how many cells are arrested at G2/M (
[0349] As a result, it was possible to confirm that, in the case of miR-122-G2U, compared with the control NT-6pi, there was no difference in number of cells at G0/G1, which is a cell cycle arrest state, but in the case of miR-122-G2,3U siRNA and miR-122-G2,7U siRNA, when a non-canonical G:A wobble seed target site was regulated in addition to G2, the proportion of the cells in the cell cycle arrest state (G0/G1) increased (
[0350] Based on the above, it was confirmed that cell cycle arrest can be increased through the regulation of non-canonical G:A wobble seed targets mediated by miR-122-G2,3U siRNA and miR-122-G2,7U siRNA.
EXAMPLE 20
Observation of Induction of Cell Cycle Arrest Induced by Regulation of Non-Canonical G:A Wobble Seed Targets Mediated by let-7a-G2U and let-7a-G2,4U
[0351] As a representative miRNA having a tumor suppressor function, there is the let-7 family. Since a function of regulating the development process of C. elegans has been reported (Nature, 2000, 403(6772): 901-906), and expression suppressed in various cancer cells (Cancer Res., 2004, 64(11): 3753-3756) and an anticancer function through the regulation of tumorigenesis (Cell, 2005, 120(5): 635-647; Genes Dev. 2007, 21(9):1025-1030) have been reported, based on this, the let-7 family has been studied as significant genes having potential for cancer diagnosis and development of an anticancer agent. Accordingly, the present inventors conducted an experiment to examine the biological function induced by the suppression of the non-canonical G:A wobble seed target of let-7.
[0352] The seed sequence from the 1.sup.st to 9.sup.th bases from the 5′ end of let-7a is 5′-UGA GGU AGU-3′, Gs capable of making G:A wobble base pairing are present in positions 2,4, 5, and 8. The inventors focused on G2 and G4, and thus conducted an experiment in the same manner as in Example 19 above, modified them into miRNA-GU to synthesize a sequence, transfected the sequence into a liver cancer cell line, that is, HepG2 cells, and then examined how a cell cycle is affected. The base sequences for the let-7 synthesized and used in the experiment are as follows: (let-7a: 5′-U GAG GUA GUA GGU UGU AUA G UU-3′; let-7a-G2U: 5′-U UAG GUA GUA GGU UGU AUA G UU-3′; let-7a-G4U: 5′-U GAU GUA GUA GGU UGU AUA G UU-3′; let-7a-G2,4U: 5′-U UAU GUA GUA GGU UGU AUA G UU-3′).
[0353] As a result, it was observed that, in the case of let-7a, compared with the control NT-6pi, there was no significant difference, but let-7a-G2U and let-7a-G2,4U increased cell cycle arrest at G0/G1, and let-7-G4U even increased the amount of cells at G2/M to make the cell cycle faster (
[0354] According to the example, let-7 serves to promote cancer cell cycle arrest by suppressing the gene of a non-canonical GA seed site at G2 based on the 5′ end, and it can be seen that, more preferably, when both G2 and G4 are involved in G:A wobble pairing, its effect is the largest. In addition, as the non-canonical GA seed site through G4 based on the 5′ end of let-7 promotes the progression of the cell cycle of cancer cells, it is considered that the proliferation of cancer cells will be induced.
EXAMPLE 21
Observation that Dedifferentiation Induced by Suppression of Non-Canonical G:A Wobble Seed Target Mediated is Promoted by miR-302-4GU and miR-372-4GU, using OCT4 Promoter Reporter
[0355] Differentiated cells may acquire differentiation ability again by artificial expression of several transcription factors, and cells finally acquiring pluripotent differentiation ability again, such as embryonic cells, are referred to as induced pluripotent stem cells. It was reported that this technique can produce inducible pluripotent stem cells (iPSs) having differentiation diversity such as stem cells by transfecting four factors (Oct3/4, Sox2, c-Myc, Klf4) into mouse embryonic fibroblasts (MEM) (Cell, 2006, 126 (4): 663-676). Similarly, it has been reported that iPSs can be made by delivering four factors (OCT4, SOX2, NANOG, and LIN28) into human somatic cells (Science, 2007, 318(5858): 1917-1920). This is a field with very high applicability in that, through dedifferentiation, the possibility of regeneration that allows pluripotent differentiation can be created by any cells, including human cells, and after the first finding, there has been a consistent effort to increase iPS production efficiency. As part of this, as one of the methods for efficiently inducing pluripotency with MiRNA-induced pluripotent stem cells (mirPSs) reported, a function of dedifferentiating cells with miRNA such as miR-302a or miR-372 alone or in combination with four factors (Oct3/4, Sox2, c-Myc and Klf4) was reported (RNA, 2008 14(10): 2115-2124; Nat Biotechnol. 2011, 29 (5): 443-448). Therefore, the inventors conducted an experiment to examine whether the inhibition of the expression of non-canonical G:A wobble seed targets of miR-302a and miR-372 can promote a process of dedifferentiating cells.
[0356] First, to monitor pluripotency induction, a Oct4 promoter reported to be activated in stem cells and a GFP-containing plasmid expressed depending on the promoter were purchased (Addgene #21319) (
[0357] First, the experiment was conducted by delivering an Oct4 expression reporter vector (pOct4:GFP) into HeLa cells using a Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's protocol, synthesizing a guide strand and a passenger strand according to each of human miR-302 and miR-372 sequences in Bionia as described in the above example to prepare a duplex, and sequentially delivering the duplexes at 50 nM using an RNAiMAX reagent (Invitrogen). In addition, siRNAs specific for the non-canonical G:A wobble seed targets of miR-302 and miR-372 were prepared by substituting a G base with U in the seed in the same manner as in the above example, and then delivered into cells at 50 nM. The base sequences of the nucleic acids used herein are as follows: (miR-302: 5′-UAAGUGCUUCCAUGUUUUGGUGA-3′; miR-302-4GU: 5′-UAAUUGCUUCCAUGUUUUGGUGA-3′; miR-302 passenger strand: 5′-ACUUAAACGUGGAUGUACUUGCU-3′; miR-372: 5′-AAAGUGCUGCGACAUUUGAGCGU-3′; miR-372-G4U: 5′-AAAUUGCUGCGACAUUUGAGCGU-3′, and miR-372 passenger strand: 5′-CCUCAAAUGUGGAGCACUAUUCU-3′). HeLa cells into which a reporter vector and RNA were transfected were incubated in Dulbecco's modified Eagle's medium (Invitrogen) supplemented with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin for 14 days, and during transfection, incubated in an antibiotic-free complete medium.
[0358] As a result, in the experimental group into which miR-302 or miR-372 was delivered independently, compared with the control, colony-forming cell growth was shown, but GFP expression showing the activity of the Oct4 promoter was not observed until the period of 14-day culture. However, in the experimental groups into which siRNAs in which the non-canonical G:A wobble seed targets of miR-302 and miR-372 are suppressed were transfected, GFP expression exhibiting colony-forming cell growth as well as the activity of the Oct4 promoter was observed (
[0359] According to this example, it was possible to observe that miR-372-G4U and miR-302a-G4U may promote the dedifferentiation of cells more efficiently than when the induction of the dedifferentiation of cells is caused by expressing conventional miR-372 and miR-302 alone, and therefore, it was able to be seen that the miR-372-G4U and the miR-302a-G4U may be used as materials for promoting the dedifferentiation of cells.
EXAMPLE 22
Confirmation of Phenomenon in Which 2′ OMe Modification at 6.SUP.th .Base Based on 5′ End Does Not Suppress Non-Canonical Nucleation Bulge Target of Corresponding RNA Interference Derivative
[0360] In the above-described example, a non-canonical nucleation bulge target had a novel biological function completely different from that of a canonical seed target, and thus miRNA-BS, which is an RNA interference-inducing nucleic acid, regulating only the inhibition of the expression of a non-canonical nucleation bulge target was invented, and its function was then confirmed. However, in the case of miRNA-BS, it was possible to confirm that, in order to complementarily arrange a non-canonical nucleation bulge site of the conventional miRNA, its seed sequence should be modified, but a novel nucleation bulge site may be generated again through the modified seed sequence. Accordingly, it is known that there is a need for technology in which an RNA interference-induced nucleic acid such as miRNA-BS recognizes only a canonical seed sequence, but does not recognize a non-canonical nucleation bulge site. Therefore, the inventors conducted an experiment to reduce the pairing strength at position 6, focusing on that the base in position 6 based on the 5′ end is important in the seed sequence for non-canonical nucleation bulge pairing, and the degree of non-canonical nucleation bulge pairing may vary depending on the pairing strength of the base in position 6. That is, a methyl group (2′ OMe) was added at the 2′ position of the ribosyl ring of the 6.sup.th nucleotide of miR-124 to modify the miR-124, and it was observed by a luciferase reporter assay whether the modified interference-inducing nucleic acid (miR-124-6me) has the function of suppressing a non-canonical nucleation bulge target of miR-124. The 2′OMe-modified miR-124 was prepared through synthesis (Bionia), and a luciferase reporter assay was conducted in the same manner as in Example 2 to measure gene inhibitory efficiency by IC50 (
[0361] As a result, it was observed that an inhibitory effect on the non-canonical bulge target gene of miR-124 shown in Example 2 disappeared, whereas the inhibition of gene expression through the canonical seed site still exhibits an excellent inhibitory effect, exhibiting an IC50 of 0.3 nM, even when 2′ OMe was added at the 6.sup.th nucleotide (
EXAMPLE 23
Confirmation that 2′ OMe Modification at 6.SUP.th .Position Based on 5′ End Does Not Suppress Total Non-Canonical Nucleation Bulge Target mRNA in Transcriptome Through RNA-Seq Analysis
[0362] In Example 22, it was confirmed that when a 2′OM modification is applied to the 6.sup.th nucleotide of miR-1, the inhibitory effect of the expression of a (miR-1-6me) canonical seed target gene is maintained, and the inhibition of the expression of a non-canonical nucleation bulge target gene is reduced. Accordingly, to confirm whether the miR-1-6me invented by the inventors, actually reduces the function of suppressing the non-canonical bulge target gene of miR-1 in a transcriptome of a cardiomyocyte cell line (h9c2) in which all genes actually related to the miR-1 function are expressed, the miR-1 and the miR-1-6me were transfected into h9c2 cells, and then subjected to RNA-Seq analysis. The RNA-Seq analysis was performed in the same manner as in Example 7 using the 6.sup.th base based on the 5′ end modified through abasic substitution (NT-6pi) in a cel-miR-67 sequence, which is miRNA of C. elegans as an experimental control. Here, an RNA-Seq library was prepared using a SENSE Total RNA-Seq Library Kit (Lexogen), and sequenced using MiniSeq (Illumina).
[0363] Afterward, the sequence data, FASTAQ file, obtained by the analysis was mapped to a mouse genomic sequence (m6) with the TopHat2 program, an expression value (FPKM) was calculated by Cufflink and Cuffdiff programs, normalized to the result in h9c2 cells into which the control NT-6pi was transfected and then expressed as a log2 ratio (fold change) for analysis. Here, to analyze the RNA-Seq result to confirm whether the amount of mRNA of a gene with a miRNA target site is inhibited by the expression of corresponding miRNA, a gene having the canonical seed site (7mer base-paired with a sequence from the 2.sup.nd to 8.sup.th bases from the 5′ end) of miR-1 in the 3′ UTR was selected, and the profile results were comparatively analyzed with a cumulative fraction in order of corresponding miRNA expression-dependent inhibition (
[0364] According to the result of the above-described example, it was able to be seen that conventional miRNA at the transcriptome level efficiently suppressed a non-canonical bulge target gene with the conventional canonical seed target gene, but miRNA in which a 2′OMe modification was applied to the 6.sup.th nucleotide (miRNA-6me) still suppresses a canonical seed target gene, but not a non-canonical bulge target gene. Accordingly, while maintaining the original intention to specifically suppress only the non-canonical nucleation bulge site of corresponding miRNA by applying the 2′OMe modification applied to the 6.sup.th base from the 5′ end to miRNA-BS, its side effects can be minimized by completely eliminating non-canonical nucleation bulge pairing that may newly appear.
EXAMPLE 24
Identification of miRNA Binding to Non-Canonical Nucleation Bulge Site Through Ago HITS CLIP Assay
[0365] To identify miRNA to which the invention for specifically recognizing and suppressing a non-canonical nucleation bulge site by miRNA can be applied, by analyzing the result by a method of analyzing a miRNA target at the transcriptome level, that is, Ago HITS CLIP assay, miRNA binding to the non-canonical nucleation bulge site was identified. First, Ago HITS-CLIP data obtained from the cerebral cortex of a mouse 13 days after birth (p13) was sequenced in the same manner as in Example 10, confirming that the top 20 expressed miRNAs binding with Argonaute bind with the non-canonical nucleation bulge site of the target mRNA (
[0366] Expanding mouse Ago HITS-CLIP data analysis of the example, other Ago HITS-CLIP results obtained from human brain and cardiac tissues (Boudreau R L et al, 2014, Neuron, 81(2) 294-305, Spengler R M et al, 2016, Nucleic Acids Res, 44(15) 7120-7131) were analyzed in the same manner as in the above-described example, and the frequencies of the canonical seed site and non-canonical nucleation bulge site at which Ago and the corresponding miRNA interacted were calculated (human brain miRNA, Table 1; human heart miRNA, Table 2).
TABLE-US-00001 TABLE 1 Seed Bulge miRNA family Seed Seed site Bulge site site # site # let-7/98/4458/4500 GAGGUAG CUACCUC UAACCUC 2,225 803 miR-125a-5p/125b-5p/351/670/4319 CCCUGAG CUCAGGG UCCAGGG 2,117 2,014 miR-124/124ab/506 AAGGCAC GUGCCUU UGGCCUU 2,345 2,707 miR-9/9ab CUUUGGU ACCAAAG CCCAAAG 2,593 2,445 miR-29abcd AGCACCA UGGUGCU GGGUGCU 3,680 1,335 miR-103a/107/107ab GCAGCAU AUGCUGC UGGCUGC 2,853 3,094 miR-221/222/222ab/1928 GCUACAU AUGUAGC UGGUAGC 1,053 830 miR-26ab/1297/4465 UCAAGUA UACUUGA ACCUUGA 1,583 1,391 miR-15abc/16/16abc/195/322/424/497/1907 AGCAGCA UGCUGCU GCCUGCU 5,278 2,420 miR-126-3p CGUACCG CGGUACG GGGUACG 97 223 miR-30abcdef/30abe-5p/384-5p GUAAACA UGUUUAC GUUUUAC 2,169 1,509 miR-33ab/33-5p UGCAUUG CAAUGCA AAAUGCA 1,464 3,217 miR-34ac/34bc-5p/449abc/449c-5p GGCAGUG CACUGCC ACCUGCC 2,450 2,360 miR-19ab GUGCAAA UUUGCAC UUUGCAC 1,735 1,735 miR-99ab/100 ACCCGUA UACGGGU ACCGGGU 191 370 miR-17/17-5p/20ab/20b- AAAGUGC GCACUUU CAACUUU 1,951 1,946 5p/93/106ab/427/518a-3p/519d miR-27abc/27a-3p UCACAGU ACUGUGA CUUGUGA 3,054 1,546 miR-218/218a UGUGCUU AAGCACA AGGCACA 2,360 1,515 miR-22/22-3p AGCUGCC GGCAGCU GCCAGCU 2,899 2,213 miR-185/882/3473/4306/4644 GGAGAGA UCUCUCC CUUCUCC 2,466 3,044 miR-181abcd/4262 ACAUUCA UGAAUGU GAAAUGU 3,504 3,159 miR-338/338-3p CCAGCAU AUGCUGG UGGCUGG 2,605 2,940 miR-127/127-3p CGGAUCC GGAUCCG GAAUCCG 430 344 miR-101/101ab ACAGUAC GUACUGU UAACUGU 1,470 1,537 miR-149 CUGGCUC GAGCCAG AGGCCAG 2,663 2,235 miR-23abc/23b-3p UCACAUU AAUGUGA AUUGUGA 3,329 2,134 miR-324-5p GCAUCCC GGGAUGC GGGAUGC 1,175 1,175 miR-24/24ab/24-3p GGCUCAG CUGAGCC UGGAGCC 2,600 2,467 miR-33a-3p/365/365-3p AAUGCCC GGGCAUU GGGCAUU 1,222 1,222 miR-139-5p CUACAGU ACUGUAG CUUGUAG 1,228 971 miR-138/138ab GCUGGUG CACCAGC ACCCAGC 3,067 2,152 miR-143/1721/4770 GAGAUGA UCAUCUC CAAUCUC 1,944 802 miR-25/32/92abc/363/363-3p/367 AUUGCAC GUGCAAU UGGCAAU 1,274 1,504 miR-574-5p GAGUGUG CACACUC ACCACUC 1,145 936 miR-7/7ab GGAAGAC GUCUUCC UCCUUCC 1,733 2,689 miR-145 UCCAGUU AACUGGA ACCUGGA 2,720 3,330 miR-135ab/135a-5p AUGGCUU AAGCCAU AGGCCAU 2,197 1,932 miR-148ab-3p/152 CAGUGCA UGCACUG GCCACUG 2,074 2,391 miR-28-5p/708/1407/1653/3139 AGGAGCU AGCUCCU GCCUCCU 2,176 2,740 miR-130ac/301ab/301b/301b- 3p/454/721/4295/3666 AGUGCAA UUGCACU UGGCACU 1,609 1,526 miR-3132 GGGUAGA UCUACCC CUUACCC 947 776 miR-155 UAAUGCU AGCAUUA GCCAUUA 1,425 1,095 miR-485-3p UCAUACA UGUAUGA GUUAUGA 1,984 1,133 miR-132/212/212-3p AACAGUC GACUGUU ACCUGUU 1,352 1,670 miR-377 UCACACA UGUGUGA GUUGUGA 2,776 1,291 hsa-miR-9-3p UAAAGCU AGCUUUA GCCUUUA 1,731 1,339 miR-374ab UAUAAUA UAUUAUA AUUUAUA 2,134 3,280 miR-129-3p/129ab-3p/129-1-3p/129-2-3p AGCCCUU AAGGGCU AGGGGCU 1,516 1,622 hsa-miR-126-5p AUUAUUA UAAUAAU AAAUAAU 2,223 3,982 miR-425/425-5p/489 AUGACAC GUGUCAU UGGUCAU 1,405 1,634 miR-423-3p GCUCGGU ACCGAGC CCCGAGC 607 1,013 miR-144 ACAGUAU AUACUGU UAACUGU 1,946 1,537 miR-21/590-5p AGCUUAU AUAAGCU UAAAGCU 951 1,714 miR-31 GGCAAGA UCUUGCC CUUUGCC 1,392 2,396 hsa-miR-20b-3p CUGUAGU ACUACAG CUUACAG 1,660 1,484 hsa-let-7d-3p UAUACGA UCGUAUA CGGUAUA 182 108 miR-191 AACGGAA UUCCGUU UCCCGUU 439 298 miR-18ab/4735-3p AAGGUGC GCACCUU CAACCUU 1,225 1,140 miR-369-3p AUAAUAC GUAUUAU UAAUUAU 1,804 2,410 hsa-miR-5187-5p GGGAUGA UCAUCCC CAAUCCC 1,543 735 miR-382 AAGUUGU ACAACUU CAAACUU 1,709 1,699 miR-485-5p/1698/1703/1962 GAGGCUG CAGCCUC AGGCCUC 3,699 1,600 hsa-miR-136-3p AUCAUCG CGAUGAU GAAUGAU 496 1,589 miR-576-3p AGAUGUG CACAUCU ACCAUCU 1,694 1,742 miR-204/204b/211 UCCCUUU AAAGGGA AAAGGGA 2,274 2,274 miR-769-5p GAGACCU AGGUCUC GGGUCUC 1,005 973 miR-342-5p/4664-5p GGGGUGC GCACCCC CAACCCC 1,387 1,516 miR-361-5p UAUCAGA UCUGAUA CUUGAUA 1,343 1,105 miR-199ab-3p/3129-5p CAGUAGU ACUACUG CUUACUG 1,295 1,409 miR-142-3p GUAGUGU ACACUAC CAACUAC 752 1,293 miR-299-5p/3563-5p GGUUUAC GUAAACC UAAAACC 716 1,482 miR-193/193b/193a-3p ACUGGCC GGCCAGU GCCCAGU 1,631 1,709 hsa-miR-1277-5p AAUAUAU AUAUAUU UAAUAUU 3,600 2,930 miR-140/140-5p/876-3p/1244 AGUGGUU AACCACU ACCCACU 1,312 1,181 hsa-miR-30a/d/e-3p UUUCAGU ACUGAAA CUUGAAA 2,764 2,569 hsa-let-7i-3p UGCGCAA UUGCGCA UGGCGCA 237 450 miR-409-5p/409a GGUUACC GGUAACC GUUAACC 650 498 miR-379/1193-5p/3529 GGUAGAC GUCUACC UCCUACC 827 1,186 miR-136 CUCCAUU AAUGGAG AUUGGAG 2,349 1,823 miR-154/872 AGGUUAU AUAACCU UAAACCU 1,004 1,282 miR-4684-3p GUUGCAA UUGCAAC UGGCAAC 991 1,385 miR-376abd/376b-3p UCAUAGA UCUAUGA CUUAUGA 1,608 1,326 miR-323/323-3p ACAUUAC GUAAUGU UAAAUGU 1,533 3,793 miR-361-3p CCCCCAG CUGGGGG UGGGGGG 2,283 1,483 miR-335/335-5p CAAGAGC GCUCUUG CUUCUUG 1,423 1,916 miR-652 AUGGCGC GCGCCAU CGGCCAU 516 680 miR-340-5p UAUAAAG CUUUAUA UUUUAUA 1,832 4,317 miR-423a/423-5p/3184/3573-5p GAGGGGC GCCCCUC CCCCCUC 2,226 2,135 miR-371/373/371b-5p CUCAAAA UUUUGAG UUUUGAG 2,605 2,605 miR-1185/3679-5p GAGGAUA UAUCCUC AUUCCUC 932 1,432 miR-3613-3p CAAAAAA UUUUUUG UUUUUUG 4,481 4,481 miR-548abakhjiwy/548abcd-5p/559 AAAGUAA UUACUUU UAACUUU 2,709 2,289 miR-93/93a/105/106a/291a- AAGUGCU AGCACUU GCCACUU 2,008 1,208 3p/294/295/302abcde/372/373/428/519a/520 be/520acd-3p/1378/1420ac miR-339b/339-5p/3586-5p CCCUGUC GACAGGG ACCAGGG 1,261 1,348 miR-876-5p/3167 GGAUUUC GAAAUCC AAAAUCC 1,627 1,856 miR-329/329ab/362-3p ACACACC GGUGUGU GUUGUGU 1,846 1,699 hsa-miR-143-5p GUGCAGU ACUGCAC CUUGCAC 1,540 1,008 miR-582-5p UACAGUU AACUGUA ACCUGUA 1,995 1,457 miR-146ac/146b-5p GAGAACU AGUUCUC GUUUCUC 1,488 1,708 miR-380/380-3p AUGUAAU AUUACAU UUUACAU 1,709 2,532 miR-487a AUCAUAC GUAUGAU UAAUGAU 1,304 1,885 miR-499-3p/499a-3p ACAUCAC GUGAUGU UGGAUGU 1,920 2,367 miR-539/539-5p GAGAAAU AUUUCUC UUUUCUC 2,102 3,454 miR-551a CGACCCA UGGGUCG GGGGUCG 298 394 miR-142-5p AUAAAGU ACUUUAU CUUUUAU 2,158 2,700 hsa-miR-17-3p CUGCAGU ACUGCAG CUUGCAG 2,741 2,227 miR-199ab-5p CCAGUGU ACACUGG CAACUGG 1,927 1,525 miR-542-3p GUGACAG CUGUCAC UGGUCAC 1,780 1,288 miR-1277 ACGUAGA UCUACGU CUUACGU 471 326 hsa-miR-29c-5p GACCGAU AUCGGUC UCCGGUC 178 270 miR-3145-3p GAUAUUU AAAUAUC AAAUAUC 1,955 1,955 hsa-miR-106b-3p CGCACUG CAGUGCG AGGUGCG 575 717 hsa-miR-22-5p GUUCUUC GAAGAAC AAAGAAC 3,059 2,606 hsa-miR-144-5p GAUAUCA UGAUAUC GAAUAUC 986 1,136 miR-744/1716 GCGGGGC GCCCCGC CCCCCGC 1,475 1,411 hsa-miR-132-5p CCGUGGC GCCACGG CCCACGG 954 829 miR-488 UGAAAGG CCUUUCA CUUUUCA 1,880 2,812 hsa-miR-377-5p GAGGUUG CAACCUC AAACCUC 1,793 1,486 miR-501-3 p/502-3p/500/502a AUGCACC GGUGCAU GUUGCAU 922 1,025 miR-486-5p/3107 CCUGUAC GUACAGG UAACAGG 1,022 915 miR-450a/451a UUUGCGA UCGCAAA CGGCAAA 253 426 hsa-miR-let7f-2/3p,hsa-miR-1185-3p UAUACAG CUGUAUA UGGUAUA 1,683 1,045 hsa-miR-30c-3p UGGGAGA UCUCCCA CUUCCCA 2,333 2,347 miR-499-5p UAAGACU AGUCUUA GUUCUUA 1,070 1,257 miR-421 UCAACAG CUGUUGA UGGUUGA 1,892 1,227 miR-197 UCACCAC GUGGUGA UGGGUGA 2,358 1,907 miR-296-5p GGGCCCC GGGGCCC GGGGCCC 1,938 1,938 miR-561 AAAGUUU AAACUUU AAACUUU 3,082 3,082 miR-326/330/330-5p CUCUGGG CCCAGAG CCCAGAG 3,106 3,106 miR-214/761/3619-5p CAGCAGG CCUGCUG CUUGCUG 4,995 2,468 miR-612/1285/3187-5p CUGGGCA UGCCCAG GCCCCAG 3,439 3,210 miR-409-3p AAUGUUG CAACAUU AAACAUU 1,798 3,158 miR-378/422a/378bcdefhi CUGGACU AGUCCAG GUUCCAG 1,653 2,031 miR-342-3p CUCACAC GUGUGAG UGGUGAG 1,793 3,079 miR-338-5p ACAAUAU AUAUUGU UAAUUGU 2,412 2,011 miR-625 GGGGGAA UUCCCCC UCCCCCC 1,847 1,202 miR-200bc/429/548a AAUACUG CAGUAUU AGGUAUU 2,342 1,840 hsa-miR-376a-5p UAGAUUC GAAUCUA AAAUCUA 942 1,785 hsa-miR-379/411-3p AUGUAAC GUUACAU UUUACAU 1,165 2,532 miR-3126-5p GAGGGAC GUCCCUC UCCCCUC 1,254 2,186 miR-584 UAUGGUU AACCAUA ACCCAUA 1,052 622 hsa-miR-let-7a/b/f-3p UAUACAA UUGUAUA UGGUAUA 2,494 1,045 miR-411 AGUAGAC GUCUACU UCCUACU 756 1,213 miR-573/3533/3616-5p/3647-5p UGAAGUG CACUUCA ACCUUCA 1,664 1,979 miR-885-5p CCAUUAC GUAAUGG UAAAUGG 1,041 1,692 hsa-miR-99-3p AAGCUCG CGAGCUU GAAGCUU 329 1,793 miR-671-5p GGAAGCC GGCUUCC GCCUUCC 2,019 2,523 miR-876-3p GGUGGUU AACCACC ACCCACC 1,386 1,692 miR-654-3p AUGUCUG CAGACAU AGGACAU 1,927 2,025 hsa-miR-340-3p CCGUCUC GAGACGG AGGACGG 924 772 miR-450b-3p/769-3p UGGGAUC GAUCCCA AUUCCCA 1,320 1,582 miR-3614-5p CACUUGG CCAAGUG CAAAGUG 1,954 2,537 hsa-miR-124-5p GUGUUCA UGAACAC GAAACAC 1,421 1,620 miR-491-5p GUGGGGA UCCCCAC CCCCCAC 2,168 2,505 miR-589 GAGAACC GGUUCUC GUUUCUC 1,006 1,708 miR-96/507/1271 UUGGCAC GUGCCAA UGGCCAA 1,652 2,566 miR-545 CAGCAAA UUUGCUG UUUGCUG 3,335 3,335 miR-548a-3p/548ef/2285a AAAACUG CAGUUUU AGGUUUU 3,014 2,238 miR-30b-3p/3689c/3689a-3p UGGGAGG CCUCCCA CUUCCCA 3,070 2,347 miR-323-5p/1421qns GGUGGUC GACCACC ACCCACC 1,458 1,692 hsa-miR-32-3p AAUUUAG CUAAAUU UAAAAUU 1,603 4,141 miR-3942-5p/4703-5p AGCAAUA UAUUGCU AUUUGCU 1,763 2,331 miR-34b/449c/1360/2682 AGGCAGU ACUGCCU CUUGCCU 2,010 1,710 hsa-miR-23a/b-5p GGGUUCC GGAACCC GAAACCC 1,112 1,544 hsa-miR-545-5p CAGUAAA UUUACUG UUUACUG 2,206 2,206 miR-362-5p/500b AUCCUUG CAAGGAU AAAGGAU 1,699 2,110 miR-677/4420 UCACUGA UCAGUGA CAAGUGA 2,556 1,859 miR-577 AGAUAAA UUUAUCU UUUAUCU 2,323 2,323 miR-3613-5p GUUGUAC GUACAAC UAACAAC 879 912 miR-369-5p GAUCGAC GUCGAUC UCCGAUC 128 172 miR-590-3p AAUUUUA UAAAAUU AAAAAUU 4,141 4,931 miR-127-5p UGAAGCU AGCUUCA GCCUUCA 1,985 2,321 miR-150/5127 CUCCCAA UUGGGAG UGGGGAG 1,828 2,905 miR-544/544ab/544-3p UUCUGCA UGCAGAA GCCAGAA 4,175 2,248 hsa-miR-29a-5p CUGAUUU AAAUCAG AAAUCAG 2,516 2,516 miR-873 CAGGAAC GUUCCUG UUUCCUG 2,349 3,190 miR-3614-3p AGCCUUC GAAGGCU AAAGGCU 1,923 1,801 miR-186 AAAGAAU AUUCUUU UUUCUUU 2,994 6,937 miR-483-3p CACUCCU AGGAGUG GGGAGUG 1,815 1,517 hsa-miR-374a-3p UUAUCAG CUGAUAA UGGAUAA 1,388 1,599 miR-196abc AGGUAGU ACUACCU CUUACCU 1,458 1,248 hsa-miR-145-3p GAUUCCU AGGAAUC GGGAAUC 1,410 891 hsa-miR-29b-2-5p UGGUUUC GAAACCA AAAACCA 2,317 3,094 hsa-miR-221-5p CCUGGCA UGCCAGG GCCCAGG 2,418 3,446 miR-323b-3p CCAAUAC GUAUUGG UAAUUGG 1,014 1,173 hsa-miR-548as-3p AAAACCC GGGUUUU GGGUUUU 1,687 1,687 miR-616 GUCAUUG CAAUGAC AAAUGAC 1,505 1,986 miR-330-3p CAAAGCA UGCUUUG GCCUUUG 2,946 2,316 hsa-miR-7-3p AACAAAU AUUUGUU UUUUGUU 3,562 6,317 miR-4525 GGGGGAU AUCCCCC UCCCCCC 926 1,202 miR-3064-5p/3085-3p CUGGCUG CAGCCAG AGGCCAG 3,803 2,235 miR-187 CGUGUCU AGACACG GAACACG 540 534 hsa-miR-26a-3p CUAUUCU AGAAUAG GAAAUAG 1,190 1,730 miR-452/4676-3p ACUGUUU AAACAGU AAACAGU 2,317 2,317 miR-129-5p/129ab-5p UUUUUGC GCAAAAA CAAAAAA 2,534 3,710 miR-223 GUCAGUU AACUGAC ACCUGAC 1,097 1,271 miR-4755-3p GCCAGGC GCCUGGC CCCUGGC 3,108 2,922 miR-1247 CCCGUCC GGACGGG GAACGGG 718 554 miR-3129-3p AACUAAU AUUAGUU UUUAGUU 1,133 2,127 hsa-miR-335-3p UUUUCAU AUGAAAA UGGAAAA 4,804 5,029 miR-542-5p CGGGGAU AUCCCCG UCCCCCG 550 763 hsa-miR-181a-3p CCAUCGA UCGAUGG CGGAUGG 432 513 hsa-miR-186-3p CCCAAAG CUUUGGG UUUUGGG 2,255 1,981 hsa-miR-96-3p AUCAUGU ACAUGAU CAAUGAU 1,619 1,601 hsa-miR-27b-5p GAGCUUA UAAGCUC AAAGCUC 658 1,376 miR-491-3p UUAUGCA UGCAUAA GCCAUAA 1,096 770 miR-4687-3p GGCUGUU AACAGCC ACCAGCC 1,686 2,279 hsa-miR-101-5p AGUUAUC GAUAACU AUUAACU 983 1,337 hsa-let-7e-3p UAUACGG CCGUAUA CGGUAUA 158 108 miR-4772-5p GAUCAGG CCUGAUC CUUGAUC 1,197 974 miR-337-3p UCCUAUA UAUAGGA AUUAGGA 950 1,012 hsa-miR-223-5p GUGUAUU AAUACAC AUUACAC 1,015 952 hsa-miR-146a-3p CUCUGAA UUCAGAG UCCAGAG 2,590 2,417 hsa-miR-16/195-3p CAAUAUU AAUAUUG AUUAUUG 2,403 2,178 miR-941 ACCCGGC GCCGGGU CCCGGGU 579 655 miR-3677-3p UCGUGGG CCCACGA CCCACGA 622 622 hsa-miR-766-5p GGAGGAA UUCCUCC UCCCUCC 2,713 2,554 miR-299/299-3p/3563-3p AUGUGGG CCCACAU CCCACAU 1,218 1,218 miR-3140-3p GCUUUUG CAAAAGC AAAAAGC 1,704 2,794 miR-532-5p/511 AUGCCUU AAGGCAU AGGGCAU 1,676 1,375 hsa-miR-24-5p GCCUACU AGUAGGC GUUAGGC 481 333 hsa-miR-4524a-3p GAGACAG CUGUCUC UGGUCUC 2,248 1,397 miR-4778-5p AUUCUGU ACAGAAU CAAGAAU 2,383 2,138 miR-642b GACACAU AUGUGUC UGGUGUC 1,522 1,749 miR-483-5p AGACGGG CCCGUCU CCCGUCU 571 571 miR-767-5p GCACCAU AUGGUGC UGGGUGC 2,048 1,223 hsa-miR-31-3p GCUAUGC GCAUAGC CAAUAGC 460 686 miR-885-3p GGCAGCG CGCUGCC GCCUGCC 1,707 2,782 miR-4706/4749-5p GCGGGGA UCCCCGC CCCCCGC 1,057 1,411 miR-574-3p ACGCUCA UGAGCGU GAAGCGU 444 423 miR-3173-3p AAGGAGG CCUCCUU CUUCCUU 2,325 2,916 miR-2127/4728-5p GGGAGGG CCCUCCC CCCUCCC 3,357 3,357 hsa-miR-103a-2-5p GCUUCUU AAGAAGC AGGAAGC 3,805 2,616 miR-3591-3p AACACCA UGGUGUU GGGUGUU 2,405 964 miR-766 CUCCAGC GCUGGAG CUUGGAG 4,936 2,384 hsa-miR-155-3p UCCUACA UGUAGGA GUUAGGA 1,168 750 hsa-miR-625-3p ACUAUAG CUAUAGU UAAUAGU 663 1,110 hsa-miR-15b-3p GAAUCAU AUGAUUC UGGAUUC 1,454 1,683 miR-522/518e/1422p AAAUGGU ACCAUUU CCCAUUU 1,930 1,550 miR-548d-3p/548acbz AAAAACC GGUUUUU GUUUUUU 2,468 3,983 hsa-miR-452-3p UCAUCUG CAGAUGA AGGAUGA 3,420 3,113 miR-192/215 UGACCUA UAGGUCA AGGGUCA 692 989 miR-1551/4524 UAGCAGC GCUGCUA CUUGCUA 1,713 911 hsa-miR-425-3p UCGGGAA UUCCCGA UCCCCGA 474 758 miR-3126-3p AUCUGGC GCCAGAU CCCAGAU 1,430 1,956 miR-519a/519bc-3p/291b-3p/1347 AAGUGCA UGCACUU GCCACUU 1,869 1,208 miR-450b-5p UUUGCAA UUGCAAA UGGCAAA 2,408 2,581 hsa-miR-125b-2-3p CACAAGU ACUUGUG CUUUGUG 1,485 3,042 miR-2441/4436a CAGGACA UGUCCUG GUUCCUG 2,535 2,349 hsa-miR-5583-3p AAUAUGG CCAUAUU CAAUAUU 1,237 1,789 miR-139-3p GAGACGC GCGUCUC CGGUCUC 530 387 miR-324-3p/1913 CUGCCCC GGGGCAG GGGGCAG 2,454 2,454 hsa-miR-141-5p AUCUUCC GGAAGAU GAAAGAU 2,912 2,391 hsa-miR-365a/b-5p GGGACUU AAGUCCC AGGUCCC 1,025 1,007 miR-654-5p/541 GGUGGGC GCCCACC CCCCACC 2,064 3,441 hsa-miR-29b-1-5p CUGGUUU AAACCAG AAACCAG 2,401 2,401 miR-563/380-5p GGUUGAC GUCAACC UCCAACC 796 1,134 hsa-miR-16-1-3p CAGUAUU AAUACUG AUUACUG 1,869 1,645 miR-1304 UUGAGGC GCCUCAA CCCUCAA 1,448 1,515 miR-216c/1461/4684-5p UCUCUAC GUAGAGA UAAGAGA 1,268 1,571 hsa-miR-2681-5p UUUUACC GGUAAAA GUUAAAA 1,871 2,348 hsa-miR-1307-5p CGACCGG CCGGUCG CGGGUCG 142 184 miR-194 GUAACAG CUGUUAC UGGUUAC 1,384 838 miR-296-3p AGGGUUG CAACCCU AAACCCU 1,287 1,644 miR-2681 AUCAUGG CCAUGAU CAAUGAU 1,560 1,601 hsa-miR-205-3p AUUUCAG CUGAAAU UGGAAAU 2,657 3,667 miR-888 ACUCAAA UUUGAGU UUUGAGU 1,927 1,927 miR-4802-3p ACAUGGA UCCAUGU CCCAUGU 1,689 1,306 hsa-miR-708-3p AACUAGA UCUAGUU CUUAGUU 1,086 956 hsa-let-7a/g-3p UGUACAG CUGUACA UGGUACA 1,922 1,356 miR-762/4492/4498 GGGCUGG CCAGCCC CAAGCCC 3,902 1,829 hsa-miR-744-3p UGUUGCC GGCAACA GCCAACA 1,638 1,906 hsa-miR-1914-3p,hsa-miR-5194 GAGGGGU ACCCCUC CCCCCUC 1,419 2,135 hsa-miR-148b-5p AGUUCUG CAGAACU AGGAACU 2,295 2,133 miR-615-5p GGGGUCC GGACCCC GAACCCC 1,701 1,133 miR-514/514b-3p UUGACAC GUGUCAA UGGUCAA 1,075 1,388 miR-28-3p ACUAGAU AUCUAGU UCCUAGU 710 818 miR-4423-5p GUUGCCU AGGCAAC GGGCAAC 1,112 1,104 miR-550a GUGCCUG CAGGCAC AGGGCAC 1,628 1,190 hsa-miR-125b-1-3p CGGGUUA UAACCCG AAACCCG 170 314 hsa-miR-506-5p AUUCAGG CCUGAAU CUUGAAU 1,601 1,672 hsa-miR-493-5p UGUACAU AUGUACA UGGUACA 2,262 1,356 hsa-miR-1306-5p CACCUCC GGAGGUG GAAGGUG 3,462 3,178 hsa-miR-561-5p UCAAGGA UCCUUGA CCCUUGA 1,593 1,071 miR-3189-3p CCUUGGG CCCAAGG CCCAAGG 2,190 2,190 miR-675-5p/4466 GGUGCGG CCGCACC CGGCACC 808 902 hsa-miR-34a-3p AAUCAGC GCUGAUU CUUGAUU 1,287 1,531 hsa-miR-454-5p CCCUAUC GAUAGGG AUUAGGG 408 522 miR-4796-3p AAAGUGG CCACUUU CAACUUU 1,584 1,946 miR-509-5p/509-3-5p/4418 ACUGCAG CUGCAGU UGGCAGU 2,886 2,144 miR-183 AUGGCAC GUGCCAU UGGCCAU 1,564 2,389 miR-182 UUGGCAA UUGCCAA UGGCCAA 2,373 2,566 hsa-miR-19a/b-5p GUUUUGC GCAAAAC CAAAAAC 1,498 2,170 hsa-miR-212-5p CCUUGGC GCCAAGG CCCAAGG 2,504 2,190 miR-3200-5p AUCUGAG CUCAGAU UCCAGAU 1,917 2,199 miR-3065-3p CAGCACC GGUGCUG GUUGCUG 3,895 2,230 miR-4755-5p UUCCCUU AAGGGAA AGGGGAA 2,544 2,094 hsa-miR-93-3p CUGCUGA UCAGCAG CAAGCAG 2,988 2,872 miR-3130-5p/4482 ACCCAGU ACUGGGU CUUGGGU 1,133 1,064 hsa-miR-488-5p CCAGAUA UAUCUGG AUUCUGG 1,210 1,862 hsa-miR-5000-5p AGUUCAG CUGAACU UGGAACU 1,839 1,923 hsa-miR-378a-5p UCCUGAC GUCAGGA UCCAGGA 1,310 3,115 miR-300-5p/4709-3p UGAAGAG CUCUUCA UCCUUCA 2,629 2,301 miR-575/4676-5p AGCCAGU ACUGGCU CUUGGCU 1,768 1,842 hsa-miR-33a-3p AAUGUUU AAACAUU AAACAUU 3,158 3,158 miR-1307 CUCGGCG CGCCGAG GCCCGAG 730 1,265 miR-3942-3p UUCAGAU AUCUGAA UCCUGAA 2,201 2,681 miR-4677-5p UGUUCUU AAGAACA AGGAACA 3,452 2,370 miR-339-3p GAGCGCC GGCGCUC GCCGCUC 659 845 miR-548b-3p AAGAACC GGUUCUU GUUUCUU 1,221 2,708 hsa-miR-642b-5p GUUCCCU AGGGAAC GGGGAAC 1,315 1,079 miR-188-5p AUCCCUU AAGGGAU AGGGGAU 1,196 1,000 hsa-miR-652-5p AACCCUA UAGGGUU AGGGGUU 668 863 miR-2114 AGUCCCU AGGGACU GGGGACU 1,293 1,145 miR-3688-5p GUGGCAA UUGCCAC UGGCCAC 1,436 2,200 hsa-miR-15a-3p AGGCCAU AUGGCCU UGGGCCU 1,693 2,093 hsa-miR-181c-3p ACCAUCG CGAUGGU GAAUGGU 348 1,407 miR-515-3p/519e AGUGCCU AGGCACU GGGCACU 1,319 1,076 miR-2447/4646-5p CUGGGAA UUCCCAG UCCCCAG 2,541 3,052 miR-122/122a/1352 GGAGUGU ACACUCC CAACUCC 904 1,498 miR-532-3p CUCCCAC GUGGGAG UGGGGAG 2,177 2,905 miR-556-3p UAUUACC GGUAAUA GUUAAUA 1,059 1,338 hsa-miR-218-2-3p AUGGUUC GAACCAU AAACCAU 1,126 2,029 miR-643 CUUGUAU AUACAAG UAACAAG 1,234 1,168 hsa-miR-92a-2-5p GGUGGGG CCCCACC CCCCACC 3,441 3,441 miR-140-3p ACCACAG CUGUGGU UGGUGGU 2,614 2,715 miR-1245 AGUGAUC GAUCACU AUUCACU 952 1,528 hsa-miR-2115-3p AUCAGAA UUCUGAU UCCUGAU 2,231 1,919 miR-93b/512-3p/1186 AGUGCUG CAGCACU AGGCACU 2,145 1,319 miR-518bcf/518a-3p/518d-3p AAAGCGC GCGCUUU CGGCUUU 417 493 miR-3200-3p ACCUUGC GCAAGGU CAAAGGU 1,413 1,958 miR-337-5p AACGGCU AGCCGUU GCCCGUU 357 313 hsa-miR-100-3p AAGCUUG CAAGCUU AAAGCUU 1,256 2,216 miR-545/3065/3065-5p CAACAAA UUUGUUG UUUGUUG 3,158 3,158 miR-17-2-3p/4793-3p CUGCACU AGUGCAG GUUGCAG 2,233 1,734 miR-1903/4778-3p CUUCUUC GAAGAAG AAAGAAG 6,522 5,458 hsa-miR-302a-5p CUUAAAC GUUUAAG UUUUAAG 1,279 3,298 hsa-miR-183-3p UGAAUUA UAAUUCA AAAUUCA 1,730 3,049 miR-3144-5p GGGGACC GGUCCCC GUUCCCC 1,177 1,090 hsa-miR-105-3p CGGAUGU ACAUCCG CAAUCCG 633 207 miR-582-3p AACUGGU ACCAGUU CCCAGUU 1,651 1,618 miR-4662a-3p AAGAUAG CUAUCUU UAAUCUU 955 1,512 miR-3140-5p CCUGAAU AUUCAGG UUUCAGG 1,829 2,876 hsa-miR-106a-3p UGCAAUG CAUUGCA AUUUGCA 1,780 2,344 hsa-miR-135a-3p AUAGGGA UCCCUAU CCCCUAU 766 569 miR-345/345-5p CUGACUC GAGUCAG AGGUCAG 1,506 1,626 hsa-miR-196a-3p GGCAACA UGUUGCC GUUUGCC 1,687 1,546 miR-125a-3p/1554 CAGGUGA UCACCUG CAACCUG 2,062 2,184 miR-3145-5p ACUCCAA UUGGAGU UGGGAGU 1,578 1,365 miR-676 UGUCCUA UAGGACA AGGGACA 731 1,542 miR-3173-5p GCCCUGC GCAGGGC CAAGGGC 1,847 1,530 hsa-miR-148a-5p AAGUUCU AGAACUU GAAACUU 2,623 2,190 hsa-miR-5586-3p AGAGUGA UCACUCU CAACUCU 1,458 1,318 miR-615-3p CCGAGCC GGCUCGG GCCUCGG 819 1,056 miR-3688-3p AUGGAAA UUUCCAU UUUCCAU 2,409 2,409 miR-4662a-5p UAGCCAA UUGGCUA UGGGCUA 1,095 890 miR-4659ab-5p UGCCAUG CAUGGCA AUUGGCA 1,724 1,588 hsa-miR-5586-5p AUCCAGC GCUGGAU CUUGGAU 1,955 1,561 hsa-miR-514a-5p ACUCUGG CCAGAGU CAAGAGU 1,626 1,358 miR-10abc/10a-5p ACCCUGU ACAGGGU CAAGGGU 1,094 850 miR-4709-5p CAACAGU ACUGUUG CUUGUUG 1,765 1,425 hsa-miR-888-3p ACUGACA UGUCAGU GUUCAGU 1,814 1,503 miR-1785/2443/3616-3p GAGGGCA UGCCCUC GCCCCUC 2,268 2,226 miR-3127-5p UCAGGGC GCCCUGA CCCCUGA 2,111 1,778 miR-1188-3p/2467-5p GAGGCUC GAGCCUC AGGCCUC 1,628 1,600 hsa-miR-382-3p AUCAUUC GAAUGAU AAAUGAU 1,589 3,166 miR-660 ACCCAUU AAUGGGU AUUGGGU 1,046 859 hsa-miR-301a-5p CUCUGAC GUCAGAG UCCAGAG 1,581 2,417 miR-508-3p GAUUGUA UACAAUC ACCAAUC 753 704 hsa-miR-185-3p GGGGCUG CAGCCCC AGGCCCC 3,623 1,750 hsa-miR-200c-5p,hsa-miR-550a-3p GUCUUAC GUAAGAC UAAAGAC 880 1,448 miR-3605-5p GAGGAUG CAUCCUC AUUCCUC 2,176 1,432 miR-513c/514b-5p UCUCAAG CUUGAGA UUUGAGA 1,417 2,980 miR-490-3p AACCUGG CCAGGUU CAAGGUU 1,423 1,176 miR-520a-5p/525-5p/2464-3p UCCAGAG CUCUGGA UCCUGGA 2,562 3,682 miR-3144-3p UAUACCU AGGUAUA GGGUAUA 1,131 416 hsa-miR-5187-3p CUGAAUC GAUUCAG AUUUCAG 1,788 2,978 miR-3664-3p CUCAGGA UCCUGAG CCCUGAG 2,354 2,643 miR-3189-5p GCCCCAU AUGGGGC UGGGGGC 1,190 1,973 miR-4670-3p GAAGUUA UAACUUC AAACUUC 1,185 1,856 miR-105/105ab CAAAUGC GCAUUUG CAAUUUG 2,029 1,390 miR-1323/5480 CAAAACU AGUUUUG GUUUUUG 2,600 2,980 hsa-miR-135b-3p UGUAGGG CCCUACA CCCUACA 1,301 1,301 hsa-miR-5010-3p UUUGUGU ACACAAA CAACAAA 2,055 2,215 miR-493/493b GAAGGUC GACCUUC ACCCUUC 1,486 1,226 miR-3605-3p CUCCGUG CACGGAG ACCGGAG 1,003 523 miR-188-3p UCCCACA UGUGGGA GUUGGGA 2,505 1,204 hsa-miR-449c-3p UGCUAGU ACUAGCA CUUAGCA 666 994 miR-486-3p GGGGCAG CUGCCCC UGGCCCC 3,671 1,979 miR-501-5p AUCCUUU AAAGGAU AAAGGAU 2,110 2,110 miR-4761-5p CAAGGUG CACCUUG ACCCUUG 1,395 1,067 miR-3130-3p CUGCACC GGUGCAG GUUGCAG 2,529 1,734 hsa-miR-202-5p UCCUAUG CAUAGGA AUUAGGA 782 1,012 miR-629 GGGUUUA UAAACCC AAAACCC 648 1,532 miR-224 AAGUCAC GUGACUU UGGACUU 1,775 2,076 miR-202-3p GAGGUAU AUACCUC UAACCUC 967 803 miR-4772-3p CUGCAAC GUUGCAG UUUGCAG 1,734 3,221 miR-4796-5p GUCUAUA UAUAGAC AUUAGAC 644 649 hsa-miR-551b-5p AAAUCAA UUGAUUU UGGAUUU 3,263 3,018 miR-556-5p AUGAGCU AGCUCAU GCCUCAU 1,783 1,498 hsa-miR-122-3p ACGCCAU AUGGCGU UGGGCGU 472 570 hsa-miR-2116-3p CUCCCAU AUGGGAG UGGGGAG 1,677 2,905 miR-4677-3p CUGUGAG CUCACAG UCCACAG 2,060 2,243 miR-877 UAGAGGA UCCUCUA CCCUCUA 1,217 886 hsa-miR-200a/b-5p AUCUUAC GUAAGAU UAAAGAU 1,426 2,480 miR-576-5p UUCUAAU AUUAGAA UUUAGAA 1,902 3,054 miR-490-5p CAUGGAU AUCCAUG UCCCAUG 1,332 1,335 hsa-miR-589-3p CAGAACA UGUUCUG GUUUCUG 2,362 2,464 hsa-miR-548a0/ax-5p GAAGUAA UUACUUC UAACUUC 1,328 1,185 miR-4786-3p GAAGCCA UGGCUUC GGGCUUC 2,438 1,736 hsa-miR-374b-3p UUAGCAG CUGCUAA UGGCUAA 1,295 1,073 hsa-miR-26b-3p CUGUUCU AGAACAG GAAACAG 2,650 2,883 miR-3158-3p AGGGCUU AAGCCCU AGGCCCU 1,924 2,078 miR-4423-3p UAGGCAC GUGCCUA UGGCCUA 947 1,191 miR-518d-5p/519bc-5p520c-5p/523b/526a UCUAGAG CUCUAGA UCCUAGA 873 1,171 miR-548aeajamx AAAAACU AGUUUUU GUUUUUU 3,406 3,983 miR-4707-3p GCCCGCC GGCGGGC GCCGGGC 833 1,405 hsa-miR-138-2-3p CUAUUUC GAAAUAG AAAAUAG 1,730 2,619 hsa-miR-10a-3p AAAUUCG CGAAUUU GAAAUUU 312 2,899 miR-526b UCUUGAG CUCAAGA UCCAAGA 1,981 2,157 miR-3622ab-3p CACCUGA UCAGGUG CAAGGUG 2,068 2,555 hsa-miR-676-5p CUUCAAC GUUGAAG UUUGAAG 1,836 3,633 hsa-miR-873-3p GAGACUG CAGUCUC AGGUCUC 1,486 1,005 hsa-miR-424-3p AAAACGU ACGUUUU CGGUUUU 718 363 hsa-miR-20a-3p CUGCAUU AAUGCAG AUUGCAG 2,577 2,148 hsa-miR-345-3p CCCUGAA UUCAGGG UCCAGGG 1,933 2,014 hsa-miR-660-3p CCUCCUG CAGGAGG AGGGAGG 3,542 2,232 hsa-miR-5004-3p UUGGAUU AAUCCAA AUUCCAA 1,667 1,752 miR-1276 AAAGAGC GCUCUUU CUUCUUU 1,806 3,023 hsa-miR-541-5p AAGGAUU AAUCCUU AUUCCUU 1,468 2,068 miR-193a-5p GGGUCUU AAGACCC AGGACCC 1,519 1,556 hsa-miR-5682 UAGCACC GGUGCUA GUUGCUA 1,266 926 miR-2115 GCUUCCA UGGAAGC GGGAAGC 2,401 1,873 miR-3663-5p CUGGUCU AGACCAG GAACCAG 2,067 1,959 hsa-miR-222-5p UCAGUAG CUACUGA UAACUGA 1,194 1,284 hsa-miR-27a-5p GGGCUUA UAAGCCC AAAGCCC 648 1,483 miR-1618/3940-3p AGCCCGG CCGGGCU CGGGGCU 932 999 miR-4661-3p AGGAUCC GGAUCCU GAAUCCU 1,282 1,248 miR-3177-5p GUGUACA UGUACAC GUUACAC 1,144 685 hsa-miR-18b-3p GCCCUAA UUAGGGC UAAGGGC 510 612 miR-146b-3p GCCCUGU ACAGGGC CAAGGGC 1,163 1,530 hsa-miR-25-5p GGCGGAG CUCCGCC UCCCGCC 1,072 998 miR-4670-5p AGCGACC GGUCGCU GUUCGCU 287 264 hsa-miR-192-3p UGCCAAU AUUGGCA UUUGGCA 1,588 2,139 hsa-miR-548at-5p AAAGUUA UAACUUU AAACUUU 2,289 3,082 miR-659 UUGGUUC GAACCAA AAACCAA 1,459 2,761 miR-141/200a AACACUG CAGUGUU AGGUGUU 2,525 1,651 hsa-miR-187-5p GCUACAA UUGUAGC UGGUAGC 1,062 830 miR-1745/3194-3p GCUCUGC GCAGAGC CAAGAGC 2,345 1,790 hsa-miR-193b-5p GGGGUUU AAACCCC AAACCCC 1,133 1,133 hsa-miR-182-3p GGUUCUA UAGAACC AGGAACC 632 1,391 miR-298/2347/2467-3p GCAGAGG CCUCUGC CUUCUGC 3,079 2,700 hsa-miR-130b-5p CUCUUUC GAAAGAG AAAAGAG 3,037 3,023 miR-2319b/3664-5p ACUCUGU ACAGAGU CAAGAGU 1,740 1,358 hsa-miR-18a-3p CUGCCCU AGGGCAG GGGGCAG 2,147 2,454 miR-3064-3p UGCCACA UGUGGCA GUUGGCA 2,551 1,379 miR-4746-3p GCGGUGC GCACCGC CAACCGC 672 553 miR-1893/2277-5p GCGCGGG CCCGCGC CCCGCGC 903 903 miR-3619-3p GGACCAU AUGGUCC UGGGUCC 819 1,045 hsa-miR-138-1-3p CUACUUC GAAGUAG AAAGUAG 1,322 1,623 hsa-miR-92a-1-5p GGUUGGG CCCAACC CCCAACC 1,397 1,397 miR-1954/3158-5p CUGCAGA UCUGCAG CUUGCAG 3,361 2,227 miR-4728-3p AUGCUGA UCAGCAU CAAGCAU 2,248 1,225 miR-3127-3p CCCCUUC GAAGGGG AAAGGGG 1,921 1,436 miR-671-3p CCGGUUC GAACCGG AAACCGG 486 315 miR-2313/3944-5p GUGCAGC GCUGCAC CUUGCAC 2,106 1,008 miR-4661-5p ACUAGCU AGCUAGU GCCUAGU 697 522 miR-509ab/509-3p GAUUGGU ACCAAUC CCCAAUC 704 664 hsa-miR-194-3p CAGUGGG CCCACUG CCCACUG 2,033 2,033 hsa-miR-211-3p CAGGGAC GUCCCUG UCCCCUG 2,263 2,087 miR-1843-5p/4802-5p AUGGAGG CCUCCAU CUUCCAU 1,752 1,757 miR-34b AAUCACU AGUGAUU GUUGAUU 1,976 1,351 hsa-miR-2114-3p GAGCCUC GAGGCUC AGGGCUC 1,445 1,230 hsa-miR-877-3p CCUCUUC GAAGAGG AAAGAGG 4,459 2,689 hsa-miR-218-1-3p UGGUUCC GGAACCA GAAACCA 1,524 2,317 miR-3157-5p UCAGCCA UGGCUGA GGGCUGA 2,470 1,581 miR-502-5p UCCUUGC GCAAGGA CAAAGGA 1,949 2,478 miR-500a AAUCCUU AAGGAUU AGGGAUU 1,913 1,178 miR-3194-5p GCCAGCC GGCUGGC GCCUGGC 2,155 3,108 hsa-miR-1304-3p CUCACUG CAGUGAG AGGUGAG 2,778 4,507 hsa-miR-5004-5p GAGGACA UGUCCUC GUUCCUC 1,991 1,405 miR-453/323b-5p GGUUGUC GACAACC ACCAACC 1,143 1,297 miR-548g AAACUGU ACAGUUU CAAGUUU 2,442 2,106 hsa-miR-92b-5p GGGACGG CCGUCCC CGGUCCC 665 541 miR-523 AACGCGC GCGCGUU CGGCGUU 120 164 hsa-miR-584-3p CAGUUCC GGAACUG GAAACUG 2,178 2,803 miR-205/205ab CCUUCAU AUGAAGG UGGAAGG 2,802 2,738 miR-4793-5p CAUCCUG CAGGAUG AGGGAUG 2,588 1,505 hsa-miR-363-5p GGGUGGA UCCACCC CCCACCC 1,794 3,038 hsa-miR-214-5p GCCUGUC GACAGGC ACCAGGC 1,129 1,458 miR-4786-5p GAGACCA UGGUCUC GGGUCUC 1,397 973 miR-3180-5p UUCCAGA UCUGGAA CUUGGAA 2,823 2,446 miR-767-3p CUGCUCA UGAGCAG GAAGCAG 2,665 3,889 hsa-miR-659-5p GGACCUU AAGGUCC AGGGUCC 853 971 miR-1404/2110 UGGGGAA UUCCCCA UCCCCCA 2,463 2,334 hsa-miR-548at-3p AAAACCG CGGUUUU GGGUUUU 363 1,687 miR-3944-3p UCGGGCU AGCCCGA GCCCCGA 770 788 miR-3157-3p UGCCCUA UAGGGCA AGGGGCA 585 1,346 hsa-miR-449b-3p AGCCACA UGUGGCU GUUGGCU 2,741 1,225 miR-3677-5p AGUGGCC GGCCACU GCCCACU 1,352 1,219 hsa-miR-191-3p CUGCGCU AGCGCAG GCCGCAG 733 1,254 miR-512-5p ACUCAGC GCUGAGU CUUGAGU 1,449 1,140 miR-1346/3940-5p/4507 UGGGUUG CAACCCA AAACCCA 1,275 1,842 miR-4746-5p CGGUCCC GGGACCG GGGACCG 671 671 miR-760-3p/1842 GGCUCUG CAGAGCC AGGAGCC 2,769 2,357 miR-3622a-5p AGGCACG CGUGCCU GUUGCCU 831 1,304 miR-3939 ACGCGCA UGCGCGU GCCGCGU 277 379 hsa-miR-181a-2-3p CCACUGA UCAGUGG CAAGUGG 2,244 1,847 miR-508-5p/509-5p ACUCCAG CUGGAGU UGGGAGU 2,469 1,365 hsa-miR-500a-3p UGCACCU AGGUGCA GGGUGCA 1,681 948 miR-4761-3p AGGGCAU AUGCCCU UGGCCCU 1,622 2,421 miR-1607/1777b/3180-3p/3196 GGGGCGG CCGCCCC CGGCCCC 1,569 1,542 miR-1914 CCUGUGC GCACAGG CAACAGG 1,697 1,589 miR-513a-5p UCACAGG CCUGUGA CUUGUGA 2,415 1,546 hsa-miR-196b-3p CGACAGC GCUGUCG CUUGUCG 684 244 hsa-miR-675-3p UGUAUGC GCAUACA CAAUACA 764 957 miR-2116 GUUCUUA UAAGAAC AAAGAAC 1,137 2,606 hsa-miR-548aj/g/x-5p GCAAAAG CUUUUGC UUUUUGC 1,945 2,486 miR-4687-5p AGCCCUC GAGGGCU AGGGGCU 1,448 1,622 miR-518a-5p/527 UGCAAAG CUUUGCA UUUUGCA 2,349 3,094 miR-4659ab-3p UUCUUCU AGAAGAA GAAAGAA 7,178 4,560 hsa-miR-5001-3p UCUGCCU AGGCAGA GGGCAGA 2,729 1,972 hsa-miR-1247-3p CCCGGGA UCCCGGG CCCCGGG 993 1,380 miR-2890/4707-5p CCCCGGC GCCGGGG CCCGGGG 1,307 1,275 hsa-miR-150-3p UGGUACA UGUACCA GUUACCA 1,539 1,064 miR-513a-3p AAAUUUC GAAAUUU AAAAUUU 2,899 4,273 hsa-miR-629-3p UUCUCCC GGGAGAA GGGAGAA 2,796 2,796 miR-548aaf AAAACCA UGGUUUU GGGUUUU 3,406 1,687 miR-2277-3p GACAGCG CGCUGUC GCCUGUC 753 1,529 hsa-miR-518c-5p CUCUGGA UCCAGAG CCCAGAG 2,417 3,106 miR-3547/3663-3p GAGCACC GGUGCUC GUUGCUC 1,534 884 miR-548k AAAGUAC GUACUUU UAACUUU 1,720 2,289 miR-34bc-3p AUCACUA UAGUGAU AGGUGAU 1,347 1,800 miR-518ef AAGCGCU AGCGCUU GCCGCUU 548 529 miR-3187-3p UGGCCAU AUGGCCA UGGGCCA 1,995 1,886 miR-4749-3p GCCCCUC GAGGGGC AGGGGGC 1,774 1,332 miR-1306/1306-3p CGUUGGC GCCAACG CCCAACG 592 516 miR-3177-3p GCACGGC GCCGUGC CCCGUGC 857 778 hsa-miR-548av-3p AAACUGC GCAGUUU CAAGUUU 2,136 2,106
TABLE-US-00002 TABLE 2 Seed Bulge miRNA family Seed Seed site Bulge site site # site # let-7/98/4458/4500 GAGGUAG CUACCUC UAACCUC 2,240 675 miR-1ab/206/613 GGAAUGU ACAUUCC CAAUUCC 1,988 841 miR-27abc/27a-3p UCACAGU ACUGUGA CUUGUGA 2,787 1,052 miR-143/1721/4770 GAGAUGA UCAUCUC CAAUCUC 1,889 586 miR-126-3p CGUACCG CGGUACG GGGUACG 87 132 miR-30a bcdef/30a be-5p/384-5p GUAAACA UGUUUAC GUUUUAC 2,334 1,157 miR-125a-5p/125b-5p/351/670/4319 CCCUGAG CUCAGGG UCCAGGG 1,531 1,331 miR-23abc/23b-3p UCACAUU AAUGUGA AUUGUGA 2,516 1,385 miR-133abc UUGGUCC GGACCAA GAACCAA 1,781 1,224 miR-15abc/16/16abc/195/322/424/497/1907 AGCAGCA UGCUGCU GCCUGCU 4,434 1,730 miR-29abcd AGCACCA UGGUGCU GGGUGCU 3,104 869 miR-499-5p UAAGACU AGUCUUA GUUCUUA 858 866 miR-99ab/100 ACCCGUA UACGGGU ACCGGGU 199 215 miR-26ab/1297/4465 UCAAGUA UACUUGA ACCUUGA 1,500 1,045 miR-21/590-5p AGCUUAU AUAAGCU UAAAGCU 715 1,218 miR-208ab/208ab-3p UAAGACG CGUCUUA GUUCUUA 257 866 miR-22/22-3p AGCUGCC GGCAGCU GCCAGCU 2,314 1,623 miR-221/222/222ab/1928 GCUACAU AUGUAGC UGGUAGC 769 576 miR-181abcd/4262 ACAUUCA UGAAUGU GAAAUGU 2,452 1,976 miR-24/24ab/24-3p GGCUCAG CUGAGCC UGGAGCC 2,063 1,763 miR-25/32/92abc/363/363-3p/367 AUUGCAC GUGCAAU UGGCAAU 1,138 1,097 miR-199ab-3p/3129-5p CAGUAGU ACUACUG CUUACUG 1,041 1,020 miR-103a/107/107ab GCAGCAU AUGCUGC UGGCUGC 2,381 2,029 miR-148ab-3p/152 CAGUGCA UGCACUG GCCACUG 1,531 1,707 miR-199ab-5p CCAGUGU ACACUGG CAACUGG 1,414 1,071 miR-28-5p/708/1407/1653/3139 AGGAGCU AGCUCCU GCCUCCU 1,530 1,754 miR-374ab UAUAAUA UAUUAUA AUUUAUA 1,538 2,233 miR-19ab GUGCAAA UUUGCAC UUUGCAC 1,522 1,522 miR-126-5p AUUAUUA UAAUAAU AAAUAAU 1,540 2,731 miR-145 UCCAGUU AACUGGA ACCUGGA 2,058 2,353 miR-193/193b/193a-3p ACUGGCC GGCCAGU GCCCAGU 1,146 1,237 miR-423a/423-5p/3184/3573-5p GAGGGGC GCCCCUC CCCCCUC 1,532 1,468 miR-33a-3p/365/365-3p AAUGCCC GGGCAUU GGGCAUU 922 922 miR-17/17-5p/20ab/20b- AAAGUGC GCACUUU CAACUUU 1,507 1,298 5p/93/106ab/427/518a-3p/519d miR-101/101ab ACAGUAC GUACUGU UAACUGU 1,086 1,106 miR-499-3p/499a-3p ACAUCAC GUGAUGU UGGAUGU 1,337 1,511 miR-214/761/3619-5p CAGCAGG CCUGCUG CUUGCUG 3,842 1,626 miR-34ac/34bc-5p/449abc/449c-5p GGCAGUG CACUGCC ACCUGCC 1,563 1,526 miR-185/882/3473/4306/4644 GGAGAGA UCUCUCC CUUCUCC 1,695 2,052 miR-378/422a/378bcdefhi CUGGACU AGUCCAG GUUCCAG 1,028 1,168 miR-218/218a UGUGCUU AAGCACA AGGCACA 1,578 970 miR-139-5p CUACAGU ACUGUAG CUUGUAG 932 655 miR-130ac/301ab/301b/301b- AGUGCAA UUGCACU UGGCACU 1,253 1,100 3p/454/721/4295/3666 miR-224 AAGUCAC GUGACUU UGGACUU 1,161 1,481 miR-30e-3p, miR-30d-3p, miR-30a-3p UUUCAGU ACUGAAA CUUGAAA 2,001 2,071 miR-146ac/146b-5p GAGAACU AGUUCUC GUUUCUC 969 1,180 miR-652 AUGGCGC GCGCCAU CGGCCAU 386 396 miR-1307 CUCGGCG CGCCGAG GCCCGAG 516 778 miR-210 UGUGCGU ACGCACA CGGCACA 356 310 miR-22-5p GUUCUUC GAAGAAC AAAGAAC 1,863 1,757 miR-338/338-3p CCAGCAU AUGCUGG UGGCUGG 2,173 1,923 miR-324-5p GCAUCCC GGGAUGC GGGAUGC 767 767 miR-128/128ab CACAGUG CACUGUG ACCUGUG 2,113 1,721 miR-191 AACGGAA UUCCGUU UCCCGUU 379 248 miR-423-3p GCUCGGU ACCGAGC CCCGAGC 419 675 miR-425/425-5p/489 AUGACAC GUGUCAU UGGUCAU 949 1,110 miR-378a-5p UCCUGAC GUCAGGA UCCAGGA 755 1,946 miR-127/127-3p CGGAUCC GGAUCCG GAAUCCG 233 216 miR-142-3p GUAGUGU ACACUAC CAACUAC 609 1,059 miR-208b-5p, miR-208a-5p AGCUUUU AAAAGCU AAAAGCU 1,680 1,680 miR-486-5p/3107 CCUGUAC GUACAGG UAACAGG 759 645 let-7d-3p UAUACGA UCGUAUA CGGUAUA 114 94 miR-33ab/33-5p UGCAUUG CAAUGCA AAAUGCA 1,075 2,362 miR-124/124ab/506 AAGGCAC GUGCCUU UGGCCUU 1,350 1,469 miR-342-3p CUCACAC GUGUGAG UGGUGAG 1,189 1,910 miR-574-5p GAGUGUG CACACUC ACCACUC 910 619 miR-18ab/4735-3p AAGGUGC GCACCUU CAACCUU 943 881 miR-190/190ab GAUAUGU ACAUAUC CAAUAUC 601 775 miR-150/5127 CUCCCAA UUGGGAG UGGGGAG 1,055 1,471 miR-223 GUCAGUU AACUGAC ACCUGAC 840 977 miR-144 ACAGUAU AUACUGU UAACUGU 1,549 1,106 miR-10abc/10a-5p ACCCUGU ACAGGGU CAAGGGU 690 541 miR-143-5p GUGCAGU ACUGCAC CUUGCAC 974 746 miR-149 CUGGCUC GAGCCAG AGGCCAG 1,861 1,451 miR-1296 UAGGGCC GGCCCUA GCCCCUA 429 432 let-7i-3p UGCGCAA UUGCGCA UGGCGCA 199 312 miR-17-3p CUGCAGU ACUGCAG CUUGCAG 1,827 1,391 miR-450a/451a UUUGCGA UCGCAAA CGGCAAA 202 281 miR-193a-5p GGGUCUU AAGACCC AGGACCC 909 970 miR-9/9ab CUUUGGU ACCAAAG CCCAAAG 2,035 1,496 miR-1307-5p CGACCGG CCGGUCG CGGGUCG 97 117 miR-132/212/212-3p AACAGUC GACUGUU ACCUGUU 856 1,211 miR-197 UCACCAC GUGGUGA UGGGUGA 1,444 1,110 miR983p, let7f-1-3p, let-7b-3p, let-7a-3p UAUACAA UUGUAUA UGGUAUA 1,849 759 miR-219-5p/508/508-3p/4782-3p GAUUGUC GACAAUC ACCAAUC 574 679 miR-24-2-5p, miR-24-1-5p GCCUACU AGUAGGC GUUAGGC 325 196 miR-144-5p GAUAUCA UGAUAUC GAAUAUC 734 754 miR-28-3p ACUAGAU AUCUAGU UCCUAGU 475 610 miR-140/140-5p/876-3p/1244 AGUGGUU AACCACU ACCCACU 945 803 miR-133a-5p GCUGGUA UACCAGC ACCCAGC 1,081 1,375 miR-598/598-3p ACGUCAU AUGACGU UGGACGU 318 566 miR-490-3p AACCUGG CCAGGUU CAAGGUU 924 768 miR-29c-5p GACCGAU AUCGGUC UCCGGUC 94 229 miR-361-5p UAUCAGA UCUGAUA CUUGAUA 862 818 miR-106b-3p CGCACUG CAGUGCG AGGUGCG 439 397 miR-93/93a/105/106a/291a- AAGUGCU AGCACUU GCCACUU 1,488 917 3p/294/295/302a bcde/372/373/428/519a/ 520be/520acd-3p/1378/1420ac miR-574-3p ACGCUCA UGAGCGU GAAGCGU 417 255 miR-421 UCAACAG CUGUUGA UGGUUGA 1,422 876 miR-99b-3p, miR-99a-3p AAGCUCG CGAGCUU GAAGCUU 278 1,241 miR-186 AAAGAAU AUUCUUU UUUCUUU 2,090 4,426 miR-181a-2-3p CCACUGA UCAGUGG CAAGUGG 1,516 1,105 miR-1404/2110 UGGGGAA UUCCCCA UCCCCCA 1,607 1,743 miR-135ab/135a-5p AUGGCUU AAGCCAU AGGCCAU 1,459 1,241 miR-362-5p/500b AUCCUUG CAAGGAU AAAGGAU 1,159 1,206 miR-153 UGCAUAG CUAUGCA UAAUGCA 818 999 miR-377 UCACACA UGUGUGA GUUGUGA 1,954 910 miR-550a GUGCCUG CAGGCAC AGGGCAC 1,047 736 miR-369-5p GAUCGAC GUCGAUC UCCGAUC 92 97 miR-140-3p ACCACAG CUGUGGU UGGUGGU 1,756 1,913 miR-6827-3p, miR-340-3p CCGUCUC GAGACGG AGGACGG 538 450 miR-214-5p GCCUGUC GACAGGC ACCAGGC 737 1,076 miR-27b-5p GAGCUUA UAAGCUC AAAGCUC 414 921 miR-296-5p GGGCCCC GGGGCCC GGGGCCC 1,212 1,212 miR-154/872 AGGUUAU AUAACCU UAAACCU 707 982 miR-204/204b/211 UCCCUUU AAAGGGA AAAGGGA 1,254 1,254 miR-145-3p GAUUCCU AGGAAUC GGGAAUC 858 601 miR-20a-3p CUGCAUU AAUGCAG AUUGCAG 1,775 1,369 miR-483-3p CACUCCU AGGAGUG GGGAGUG 1,019 932 miR-155 UAAUGCU AGCAUUA GCCAUUA 1,024 773 miR-23b-5p, miR-23a-5p GGGUUCC GGAACCC GAAACCC 812 977 miR-1277 ACGUAGA UCUACGU CUUACGU 326 194 miR-664/664b AUUCAUU AAUGAAU AUUGAAU 1,850 1,189 miR-887 UGAACGG CCGUUCA CGGUUCA 218 213 miR-495/1192 AACAAAC GUUUGUU UUUUGUU 1,700 4,230 miR-369-3p AUAAUAC GUAUUAU UAAUUAU 1,234 1,440 miR-361-3p CCCCCAG CUGGGGG UGGGGGG 1,372 836 miR-92b-5p GGGACGG CCGUCCC CGGUCCC 478 422 miR-339b/339-5p/3586-5p CCCUGUC GACAGGG ACCAGGG 719 957 miR-340-5p UAUAAAG CUUUAUA UUUUAUA 1,351 2,941 miR-136-3p AUCAUCG CGAUGAU GAAUGAU 302 910 miR-503 AGCAGCG CGCUGCU GCCUGCU 1,093 1,730 miR-1287 GCUGGAU AUCCAGC UCCCAGC 1,074 1,958 miR-654-3p AUGUCUG CAGACAU AGGACAU 1,386 1,351 miR-532-5p/511 AUGCCUU AAGGCAU AGGGCAU 1,058 908 miR-1-5p CAUACUU AAGUAUG AGGUAUG 1,424 1,229 miR-493-5p UGUACAU AUGUACA UGGUACA 1,770 1,176 miR-744/1716 GCGGGGC GCCCCGC CCCCCGC 1,134 952 miR-3591-3p AACACCA UGGUGUU GGGUGUU 1,492 632 miR-30b-3p/3689c/3689a-3p UGGGAGG CCUCCCA CUUCCCA 2,081 1,684 miR-329/329ab/362-3p ACACACC GGUGUGU GUUGUGU 988 1,103 miR-4524a-3p GAGACAG CUGUCUC UGGUCUC 1,533 1,010 miR-376c/741-5p ACAUAGA UCUAUGU CUUAUGU 932 992 miR-29b-2-5p UGGUUUC GAAACCA AAAACCA 1,592 2,292 miR-193b-5p GGGGUUU AAACCCC AAACCCC 752 752 miR-374c/655 UAAUACA UGUAUUA GUUAUUA 1,713 878 miR-335/335-5p CAAGAGC GCUCUUG CUUCUUG 892 1,332 miR-452/4676-3p ACUGUUU AAACAGU AAACAGU 1,589 1,589 miR-96/507/1271 UUGGCAC GUGCCAA UGGCCAA 1,198 1,841 miR-494 GAAACAU AUGUUUC UGGUUUC 1,415 1,185 miR-136 CUCCAUU AAUGGAG AUUGGAG 1,329 1,057 miR-29a-5p CUGAUUU AAAUCAG AAAUCAG 1,660 1,660 miR-142-5p AUAAAGU ACUUUAU CUUUUAU 1,660 1,832 miR-501-3p/502-3p/500/502a AUGCACC GGUGCAU GUUGCAU 546 707 miR-542-5p CGGGGAU AUCCCCG UCCCCCG 346 546 miR-874 UGCCCUG CAGGGCA AGGGGCA 1,328 835 miR-660 ACCCAUU AAUGGGU AUUGGGU 649 513 miR-138/138ab GCUGGUG CACCAGC ACCCAGC 1,898 1,375 miR-7/7ab GGAAGAC GUCUUCC UCCUUCC 1,236 2,073 miR-101-5p AGUUAUC GAUAACU AUUAACU 600 907 miR-651 UUAGGAU AUCCUAA UCCCUAA 585 677 miR-339-3p GAGCGCC GGCGCUC GCCGCUC 448 516 miR-625 GGGGGAA UUCCCCC UCCCCCC 1,259 942 miR-584 UAUGGUU AACCAUA ACCCAUA 657 437 miR-551a CGACCCA UGGGUCG GGGGUCG 191 232 miR-1301/5047 UGCAGCU AGCUGCA GCCUGCA 2,276 1,750 miR-31 GGCAAGA UCUUGCC CUUUGCC 979 1,660 miR-381-5p GCGAGGU ACCUCGC CCCUCGC 296 547 miR-345/345-5p CUGACUC GAGUCAG AGGUCAG 846 950 miR-374a-3p UUAUCAG CUGAUAA UGGAUAA 874 928 miR-181a-3p CCAUCGA UCGAUGG CGGAUGG 245 294 miR-8073, miR-221-5p CCUGGCA UGCCAGG GCCCAGG 1,403 1,893 miR-500a-3p UGCACCU AGGUGCA GGGUGCA 959 647 miR-409-5p/409a GGUUACC GGUAACC GUUAACC 351 393 miR-542-3p GUGACAG CUGUCAC UGGUCAC 1,326 878 miR-491-5p GUGGGGA UCCCCAC CCCCCAC 1,514 1,749 miR-6788-5p, miR-30c-2-3p, miR-30c-1-3p UGGGAGA UCUCCCA CUUCCCA 1,484 1,684 miR-216a AAUCUCA UGAGAUU GAAGAUU 1,070 1,489 miR-194 GUAACAG CUGUUAC UGGUUAC 1,079 624 miR-941 ACCCGGC GCCGGGU CCCGGGU 403 377 miR-300/381/539-3p AUACAAG CUUGUAU UUUGUAU 1,188 2,820 miR-376a-5p UAGAUUC GAAUCUA AAAUCUA 720 1,104 miR-1306-5p CACCUCC GGAGGUG GAAGGUG 2,013 1,894 miR-1249 CGCCCUU AAGGGCG AGGGGCG 280 385 miR-485-3p UCAUACA UGUAUGA GUUAUGA 1,234 678 miR-642a UCCCUCU AGAGGGA GAAGGGA 1,083 1,205 miR-532-3p CUCCCAC GUGGGAG UGGGGAG 1,306 1,471 miR-3613-5p GUUGUAC GUACAAC UAACAAC 624 596 miR-424-3p AAAACGU ACGUUUU CGGUUUU 505 224 miR-769-5p GAGACCU AGGUCUC GGGUCUC 603 633 miR-3200-3p ACCUUGC GCAAGGU CAAAGGU 826 1,262 miR-1185-2-3p, miR-1185-1-3p, let-7f-2-3p UAUACAG CUGUAUA UGGUAUA 1,249 759 miR-486-3p GGGGCAG CUGCCCC UGGCCCC 2,110 1,257 let-7g-3p,let-7a-2-3p UGUACAG CUGUACA UGGUACA 1,502 1,176 miR-125b-1-3p CGGGUUA UAACCCG AAACCCG 112 227 let-7e-3p UAUACGG CCGUAUA CGGUAUA 152 94 miR-26b-3p CUGUUCU AGAACAG GAAACAG 1,736 1,874 miR-487a AUCAUAC GUAUGAU UAAUGAU 916 1,120 miR-132-5p CCGUGGC GCCACGG CCCACGG 602 500 miR-382 AAGUUGU ACAACUU CAAACUU 1,321 1,177 miR-579 UCAUUUG CAAAUGA AAAAUGA 1,423 3,031 miR-766 CUCCAGC GCUGGAG CUUGGAG 3,325 1,581 miR-493/493b GAAGGUC GACCUUC ACCCUUC 934 932 miR-326/330/330-5p CUCUGGG CCCAGAG CCCAGAG 1,717 1,717 miR-299-5p/3563-5p GGUUUAC GUAAACC UAAAACC 643 1,034 miR-202-5p UCCUAUG CAUAGGA AUUAGGA 493 630 miR-374b-3p UUAGCAG CUGCUAA UGGCUAA 1,062 731 miR-3605-3p CUCCGUG CACGGAG ACCGGAG 683 270 miR-210-5p GCCCCUG CAGGGGC AGGGGGC 1,060 881 miR-15a-3p AGGCCAU AUGGCCU UGGGCCU 1,079 1,332 miR-382-3p AUCAUUC GAAUGAU AAAUGAU 910 1,945 miR-93-3p CUGCUGA UCAGCAG CAAGCAG 1,960 1,934 miR-195-3p, miR-16-2-3p CAAUAUU AAUAUUG AUUAUUG 1,515 1,474 miR-187 CGUGUCU AGACACG GAACACG 334 307 miR-5010-3p UUUGUGU ACACAAA CAACAAA 1,493 1,602 miR-103a-2-5p GCUUCUU AAGAAGC AGGAAGC 2,642 1,824 miR-33a-3p AAUGUUU AAACAUU AAACAUU 2,352 2,352 miR-548a-3p/548ef/2285a AAAACUG CAGUUUU AGGUUUU 2,073 1,444 miR-490-5p CAUGGAU AUCCAUG UCCCAUG 825 986 miR-522/518e/1422p AAAUGGU ACCAUUU CCCAUUU 1,444 1,063 miR-1306/1306-3p CGUUGGC GCCAACG CCCAACG 426 385 miR-376abd/376b-3p UCAUAGA UCUAUGA CUUAUGA 1,089 914 miR-675-5p/4466 GGUGCGG CCGCACC CGGCACC 524 570 miR-677/4420 UCACUGA UCAGUGA CAAGUGA 1,670 1,276 miR-509-5p/509-3-5p/4418 ACUGCAG CUGCAGU UGGCAGU 2,130 1,444 miR-125a-3p/1554 CAGGUGA UCACCUG CAACCUG 1,482 1,582 miR-200bc/429/548a AAUACUG CAGUAUU AGGUAUU 1,782 1,010 miR-130b-5p CUCUUUC GAAAGAG AAAAGAG 1,625 1,864 miR-656 AUAUUAU AUAAUAU UAAAUAU 1,435 2,879 miR-192/215 UGACCUA UAGGUCA AGGGUCA 362 639 miR-337-3p UCCUAUA UAUAGGA AUUAGGA 586 630 miR-548aaf AAAACCA UGGUUUU GGGUUUU 2,131 947 miR-139-3p GGAGACG CGUCUCC GUUCUCC 516 932 miR-337-5p AACGGCU AGCCGUU GCCCGUU 231 223 miR-589 GAGAACC GGUUCUC GUUUCUC 631 1,180 miR-3200-5p AUCUGAG CUCAGAU UCCAGAU 1,433 1,530 miR-323/323-3p ACAUUAC GUAAUGU UAAAUGU 920 2,441 miR-488 UGAAAGG CCUUUCA CUUUUCA 1,315 1,862 miR-1185/3679-5p GAGGAUA UAUCCUC AUUCCUC 593 1,283 miR-6511b-3p, miR-6511a-3p CUCACCA UGGUGAG GGGUGAG 1,910 1,229 miR-660-3p CCUCCUG CAGGAGG AGGGAGG 1,858 965 miR-203 UGAAAUG CAUUUCA AUUUUCA 1,903 2,455 miR-425-3p UCGGGAA UUCCCGA UCCCCGA 391 548 miR-433 UCAUGAU AUCAUGA UCCAUGA 1,244 1,137 miR-450b-5p UUUGCAA UUGCAAA UGGCAAA 1,666 1,652 miR-675-3p UGUAUGC GCAUACA CAAUACA 555 729 miR-1180 UUCCGGC GCCGGAA CCCGGAA 330 415 miR-342-5p/4664-5p GGGGUGC GCACCCC CAACCCC 970 1,038 miR-597/1970 GUGUCAC GUGACAC UGGACAC 800 1,168 miR-26a-2-3p, miR-26a-1-3p CUAUUCU AGAAUAG GAAAUAG 685 989 miR-338-5p ACAAUAU AUAUUGU UAAUUGU 1,765 1,291 miR-324-3p/1913 CUGCCCC GGGGCAG GGGGCAG 1,474 1,474 miR-548abakhjiwy/548abcd-5p/559 AAAGUAA UUACUUU UAACUUU 2,013 1,608 miR-518a-5p/527 UGCAAAG CUUUGCA UUUUGCA 1,746 2,241 miR-9-3p UAAAGCU AGCUUUA GCCUUUA 1,258 911 miR-299/299-3p/3563-3p AUGUGGG CCCACAU CCCACAU 906 906 miR-27a-5p GGGCUUA UAAGCCC AAAGCCC 451 939 miR-491-3p UUAUGCA UGCAUAA GCCAUAA 654 600 miR-671-5p GGAAGCC GGCUUCC GCCUUCC 1,385 1,413 miR-190a-3p UAUAUAU AUAUAUA UAAUAUA 2,181 1,454 miR-1323/5480 CAAAACU AGUUUUG GUUUUUG 1,639 1,882 miR-1537 AAACCGU ACGGUUU CGGGUUU 261 268 miR-411 AGUAGAC GUCUACU UCCUACU 559 1,008 miR-4662a-5p UAGCCAA UUGGCUA UGGGCUA 734 574 miR-411-3p, miR-379-3p AUGUAAC GUUACAU UUUACAU 800 1,934 miR-550a-3p, miR-200c-5p GUCUUAC GUAAGAC UAAAGAC 553 959 miR-141/200a AACACUG CAGUGUU AGGUGUU 1,731 1,002 miR-3613-3p CAAAAAA UUUUUUG UUUUUUG 3,104 3,104 miR-2277-3p GACAGCG CGCUGUC GCCUGUC 521 1,103 miR-545/3065/3065-5p CAACAAA UUUGUUG UUUGUUG 1,984 1,984 miR-500a AAUCCUU AAGGAUU AGGGAUU 1,056 680 miR-545-5p CAGUAAA UUUACUG UUUACUG 1,621 1,621 miR-409-3p AAUGUUG CAACAUU AAACAUU 1,478 2,352 miR-874-5p GGCCCCA UGGGGCC GGGGGCC 1,385 1,095 miR-410/344de/344b-1-3p AUAUAAC GUUAUAU UUUAUAU 963 2,815 miR-576-5p UUCUAAU AUUAGAA UUUAGAA 1,231 1,952 miR-129-5p/129ab-5p UUUUUGC GCAAAAA CAAAAAA 1,602 2,408 miR-370 CCUGCUG CAGCAGG AGGCAGG 2,014 1,252 miR-1296-3p AGUGGGG CCCCACU CCCCACU 1,356 1,356 miR-590-3p AAUUUUA UAAAAUU AAAAAUU 2,634 3,067 miR-519a/519bc-3p/291b-3p/1347 AAGUGCA UGCACUU GCCACUU 1,434 917 miR-518bcf/518a-3p/518d-3p AAAGCGC GCGCUUU CGGCUUU 286 326 miR-501-5p AUCCUUU AAAGGAU AAAGGAU 1,206 1,206 miR-629 GGGUUUA UAAACCC AAAACCC 515 1,101 miR-889 UAAUAUC GAUAUUA AUUAUUA 794 1,590 miR-450b-3p/769-3p UGGGAUC GAUCCCA AUUCCCA 820 1,347 miR-1618/3940-3p AGCCCGG CCGGGCU CGGGGCU 599 572 miR-125b-2-3p CACAAGU ACUUGUG CUUUGUG 959 2,232 miR-548d-3p/548acbz AAAAACC GGUUUUU GUUUUUU 1,511 2,520 miR-485-5p/1698/1703/1962 GAGGCUG CAGCCUC AGGCCUC 2,388 1,087 miR-127-5p UGAAGCU AGCUUCA GCCUUCA 1,476 1,469 miR-6086, miR-377-5p GAGGUUG CAACCUC AAACCUC 1,385 1,093 miR-183 AUGGCAC GUGCCAU UGGCCAU 1,045 1,385 miR-625-3p ACUAUAG CUAUAGU UAAUAGU 485 738 miR-544/544ab/544-3p UUCUGCA UGCAGAA GCCAGAA 2,624 1,422 miR-31-3p GCUAUGC GCAUAGC CAAUAGC 321 480 miR-454-5p CCCUAUC GAUAGGG AUUAGGG 195 309 miR-539/539-5p GAGAAAU AUUUCUC UUUUCUC 1,368 2,373 miR-487b-5p, miR-487a-5p UGGUUAU AUAACCA UAAACCA 751 1,077 miR-545 CAGCAAA UUUGCUG UUUGCUG 2,422 2,422 miR-4525 GGGGGAU AUCCCCC UCCCCCC 712 942 miR-1304-3p CUCACUG CAGUGAG AGGUGAG 1,694 2,510 miR-4423-5p GUUGCCU AGGCAAC GGGCAAC 638 707 miR-16-1-3p CAGUAUU AAUACUG AUUACUG 1,396 1,461 miR-652-5p AACCCUA UAGGGUU AGGGGUU 337 449 miR-219-2-3p/219-3p GAAUUGU ACAAUUC CAAAUUC 771 1,149 miR-1247 CCCGUCC GGACGGG GAACGGG 462 376 miR-181c-3p ACCAUCG CGAUGGU GAAUGGU 333 924 miR-15b-3p GAAUCAU AUGAUUC UGGAUUC 1,007 970 miR-3620 CACCCUG CAGGGUG AGGGGUG 1,059 703 miR-885-5p CCAUUAC GUAAUGG UAAAUGG 559 1,086 miR-380/380-3p AUGUAAU AUUACAU UUUACAU 1,360 1,934 miR-379/1193-5p/3529 GGUAGAC GUCUACC UCCUACC 609 886 miR-656-5p GGUUGCC GGCAACC GCCAACC 638 937 miR-188-5p AUCCCUU AAGGGAU AGGGGAU 725 591 miR-744-3p UGUUGCC GGCAACA GCCAACA 964 1,457 miR-122/122a/1352 GGAGUGU ACACUCC CAACUCC 760 995 miR-671-3p CCGGUUC GAACCGG AAACCGG 320 216 miR 7 2 3p, miR 7 1 3p AACAAAU AUUUGUU UUUUGUU 2,401 4,230 miR-518d-5p/519bc-5p520c-5p/523b/526a UCUAGAG CUCUAGA UCCUAGA 594 668 miR-34a-3p AAUCAGC GCUGAUU CUUGAUU 870 986 miR-504/4725-5p GACCCUG CAGGGUC AGGGGUC 801 417 miR-301b-5p, miR-301a-5p CUCUGAC GUCAGAG UCCAGAG 1,035 1,751 miR-654-5p/541 GGUGGGC GCCCACC CCCCACC 1,464 2,305 miR-29b-1-5p CUGGUUU AAACCAG AAACCAG 1,830 1,830 miR-371/373/371b-5p CUCAAAA UUUUGAG UUUUGAG 1,634 1,634 miR-2964/2964a-5p GAUGUCC GGACAUC GAACAUC 1,097 1,143 miR-520a-5p/525-5p/2464-3p UCCAGAG CUCUGGA UCCUGGA 1,639 2,485 miR-450a-2-3p UUGGGGA UCCCCAA CCCCCAA 1,381 1,329 miR-511 UGUCUUU AAAGACA AAAGACA 2,007 2,007 miR-676 UGUCCUA UAGGACA AGGGACA 478 1,028 miR-3130-5p/4482 ACCCAGU ACUGGGU CUUGGGU 727 663 miR-3173-5p GCCCUGC GCAGGGC CAAGGGC 1,195 1,045 miR-296-3p AGGGUUG CAACCCU AAACCCU 832 1,127 miR-708-3p AACUAGA UCUAGUU CUUAGUU 657 703 miR-182 UUGGCAA UUGCCAA UGGCCAA 1,649 1,841 miR-511-3p AUGUGUA UACACAU ACCACAU 1,072 1,010 miR-1277-5p AAUAUAU AUAUAUU UAAUAUU 2,437 2,072 miR-3064-3p UGCCACA UGUGGCA GUUGGCA 1,804 877 miR-205/205ab CCUUCAU AUGAAGG UGGAAGG 1,579 1,550 miR-323b-3p CCAAUAC GUAUUGG UAAUUGG 593 647 miR-1893/2277-5p GCGCGGG CCCGCGC CCCGCGC 560 560 miR-3157-5p UCAGCCA UGGCUGA GGGCUGA 1,734 1,060 miR-582-5p UACAGUU AACUGUA ACCUGUA 1,375 1,075 miR-1343 UCCUGGG CCCAGGA CCCAGGA 2,022 2,022 miR-365b-5p, miR-365a-5p GGGACUU AAGUCCC AGGUCCC 658 659 miR-146b-3p GCCCUGU ACAGGGC CAAGGGC 790 1,045 miR-124-5p GUGUUCA UGAACAC GAAACAC 1,018 1,146 miR-6505-3p GACUUCU AGAAGUC GAAAGUC 1,061 975 miR-526b UCUUGAG CUCAAGA UCCAAGA 1,429 1,346 miR-548aeajamx AAAAACU AGUUUUU GUUUUUU 2,155 2,520 miR-4794 CUGGCUA UAGCCAG AGGCCAG 735 1,451 miR-2114 AGUCCCU AGGGACU GGGGACU 785 726 miR-19b-2-5p, miR-19b-1-5p, miR-19a-5p GUUUUGC GCAAAAC CAAAAAC 1,020 1,590 miR-18a-3p CUGCCCU AGGGCAG GGGGCAG 1,404 1,474 miR-185-3p GGGGCUG CAGCCCC AGGCCCC 2,449 1,084 miR-433-5p ACGGUGA UCACCGU CAACCGU 453 226 let-7c-3p UGUACAA UUGUACA UGGUACA 1,747 1,176 miR-134/3118 GUGACUG CAGUCAC AGGUCAC 1,108 678 miR-548b-3p AAGAACC GGUUCUU GUUUCUU 825 1,825 miR-876-3p GGUGGUU AACCACC ACCCACC 1,020 1,264 miR-3132 GGGUAGA UCUACCC CUUACCC 630 596 miR-10a-3p AAAUUCG CGAAUUU GAAAUUU 177 1,754 miR-6741-3p CGGCUCU AGAGCCG GAAGCCG 489 638 miR-34bc-3p AUCACUA UAGUGAU AGGUGAU 842 1,094 miR-186-3p CCCAAAG CUUUGGG UUUUGGG 1,451 1,349 miR-4661-5p ACUAGCU AGCUAGU GCCUAGU 479 409 miR-370-5p AGGUCAC GUGACCU UGGACCU 1,173 1,444 miR-653-3p UCACUGG CCAGUGA CAAGUGA 1,773 1,276 miR-5001-3p UCUGCCU AGGCAGA GGGCAGA 1,606 1,222 miR-2964a-3p GAAUUGC GCAAUUC CAAAUUC 627 1,149 miR-330-3p CAAAGCA UGCUUUG GCCUUUG 2,124 1,436 miR-579-5p CGCGGUU AACCGCG ACCCGCG 114 192 miR-3064-5p/3085-3p CUGGCUG CAGCCAG AGGCCAG 2,582 1,451 miR-376c-5p, miR-376b-5p GUGGAUA UAUCCAC AUUCCAC 537 1,011 miR-3127-3p CCCCUUC GAAGGGG AAAGGGG 1,091 803 miR-483-5p AGACGGG CCCGUCU CCCGUCU 345 345 miR-129-3p/129ab-3p/129-1-3p/129-2-3p AGCCCUU AAGGGCU AGGGGCU 843 1,016 miR-196abc AGGUAGU ACUACCU CUUACCU 1,243 1,010 miR-576-3p AGAUGUG CACAUCU ACCAUCU 1,248 1,205 miR-552/3097-5p ACAGGUG CACCUGU ACCCUGU 1,448 1,073 miR-4761-5p CAAGGUG CACCUUG ACCCUUG 1,009 678 miR-1745/3194-3p GCUCUGC GCAGAGC CAAGAGC 1,601 1,220 miR-4707-3p GCCCGCC GGCGGGC GCCGGGC 742 1,085 miR-548ay-3p, miR-548at-3p AAAACCG CGGUUUU GGGUUUU 224 947 miR-34b/449c/1360/2682 AGGCAGU ACUGCCU CUUGCCU 1,215 1,209 miR-513a-5p UCACAGG CCUGUGA CUUGUGA 1,826 1,052 miR-3145-5p ACUCCAA UUGGAGU UGGGAGU 1,013 832 miR-3158-3p AGGGCUU AAGCCCU AGGCCCU 1,186 1,293 miR-556-5p AUGAGCU AGCUCAU GCCUCAU 1,199 1,040 miR-3194-5p GCCAGCC GGCUGGC GCCUGGC 1,385 1,958 miR-6734-3p CCUUCCC GGGAAGG GGGAAGG 1,402 1,402 miR-523 AACGCGC GCGCGUU CGGCGUU 94 152 miR-1910 CAGUCCU AGGACUG GGGACUG 1,213 1,071 miR-4670-5p AGCGACC GGUCGCU GUUCGCU 182 203 miR-2115 GCUUCCA UGGAAGC GGGAAGC 1,641 1,091 miR-508-5p/509-5p ACUCCAG CUGGAGU UGGGAGU 1,518 832 miR-1245 AGUGAUC GAUCACU AUUCACU 701 1,223 miR-556-3p UAUUACC GGUAAUA GUUAAUA 596 908 miR-188-3p UCCCACA UGUGGGA GUUGGGA 1,513 797 miR-651-3p AAGGAAA UUUCCUU UUUCCUU 2,819 2,819 miR-615-3p CCGAGCC GGCUCGG GCCUCGG 496 608 miR-758 UUGUGAC GUCACAA UCCACAA 663 1,125 miR-4670-3p GAAGUUA UAACUUC AAACUUC 934 1,371 miR-6874-3p, miR-148b-5p AGUUCUG CAGAACU AGGAACU 1,585 1,447 miR-4778-5p AUUCUGU ACAGAAU CAAGAAU 1,649 1,515 miR-453/323b-5p GGUUGUC GACAACC ACCAACC 768 997 miR-6735-3p GGCCUGU ACAGGCC CAAGGCC 896 1,384 miR-2116-3p CUCCCAU AUGGGAG UGGGGAG 944 1,471 miR-4677-5p UGUUCUU AAGAACA AGGAACA 2,302 1,435 miR-584-3p CAGUUCC GGAACUG GAAACUG 1,412 1,733 miR-885-3p GGCAGCG CGCUGCC GCCUGCC 975 1,819 miR-1287-3p UCUAGCC GGCUAGA GCCUAGA 338 414 miR-2127/4728-5p GGGAGGG CCCUCCC CCCUCCC 2,391 2,391 miR-93b/512-3p/1186 AGUGCUG CAGCACU AGGCACU 1,487 923 miR-32-3p AAUUUAG CUAAAUU UAAAAUU 958 2,634 miR-561-5p UCAAGGA UCCUUGA CCCUUGA 1,163 769 miR-766-5p GGAGGAA UUCCUCC UCCCUCC 1,925 1,813 miR-877 UAGAGGA UCCUCUA CCCUCUA 843 601 miR-25-5p GGCGGAG CUCCGCC UCCCGCC 734 607 miR-376a-2-5p GUAGAUU AAUCUAC AUUCUAC 610 763 miR-585 GGGCGUA UACGCCC ACCGCCC 231 454 miR-3187-3p UGGCCAU AUGGCCA UGGGCCA 1,342 1,478 miR-605 AAAUCCC GGGAUUU GGGAUUU 862 862
[0367] As a result, it was possible to confirm that miRNAs listed in the tables (Tables 1 and 2) bind with a non-canonical nucleation bulge site in a corresponding tissue. Therefore, when all of the data obtained through the Ago HITS-CLIP assay in the example was summarized, thereby identifying miRNA, and then the present invention for modifying a non-canonical nucleation bulge site to be recognized as a canonical seed site was applied to the miRNA, as shown in Table 3 below, each of a total of 426 sequences (BS sequences) consists of the 2.sup.nd to 7.sup.th nucleotides based on the 5′ end of an RNA interference nucleic acid, and the modified RNA interference nucleic acid will specifically bind to the non-canonical bulge target of the corresponding miRNA and exhibit only the corresponding function.
TABLE-US-00003 TABLE 3 BS SEQ Bulge sequence ID NOs miRNA family Seed Seed site site GAGGUU 103 let-7/98/4458/4500 GAGGUA UACCUC AACCUC CCCUGG 104 miR-125a-5p/125b-5p/351/670/4319 CCCUGA UCAGGG CCAGGG AAGGCC 105 miR-124/124ab/506 AAGGCA UGCCUU GGCCUU CUUUGG 106 miR-9/9ab CUUUGG CCAAAG CCAAAG AGCACC 107 miR-29a bcd AGCACC GGUGCU GGUGCU GCAGCC 108 miR-103a/107/107ab GCAGCA UGCUGC GGCUGC GCUACC 109 miR-221/222/222ab/1928 GCUACA UGUAGC GGUAGC UCAAGG 110 miR-26ab/1297/4465 UCAAGU ACUUGA CCUUGA AGCAGG 111 miR-15abc/16/16abc/195/322/424/497/1907 AGCAGC GCUGCU CCUGCU CGUACC 112 miR-126-3p CGUACC GGUACG GGUACG GUAAAA 113 miR-30abcdef/30abe-5p/384-5p GUAAAC GUUUAC UUUUAC UGCAUU 114 miR-33ab/33-5p UGCAUU AAUGCA AAUGCA GGCAGG 115 miR-34ac/34bc-5p/449abc/449c-5p GGCAGU ACUGCC CCUGCC GUGCAA 116 miR-19ab GUGCAA UUGCAC UUGCAC ACCCGG 117 miR-99ab/100 ACCCGU ACGGGU CCGGGU AAAGUU 118 miR-17/17-5p/20ab/20b-5p/93/106ab/427/518a- AAAGUG CACUUU AACUUU 3p/519d UCACAA 119 miR-27abc/27a-3p UCACAG CUGUGA UUGUGA UGUGCC 120 miR-218/218a UGUGCU AGCACA GGCACA AGCUGG 121 miR-22/22-3p AGCUGC GCAGCU CCAGCU GGAGAA 122 miR-185/882/3473/4306/4644 GGAGAG CUCUCC UUCUCC ACAUUU 123 miR-181abcd/4262 ACAUUC GAAUGU AAAUGU CCAGCC 124 miR-338/338-3p CCAGCA UGCUGG GGCUGG CGGAUU 125 miR-127/127-3p CGGAUC GAUCCG AAUCCG ACAGUU 126 miR-101/101ab ACAGUA UACUGU AACUGU CUGGCC 127 miR-149 CUGGCU AGCCAG GGCCAG GCAUCC 128 miR-324-5p GCAUCC GGAUGC GGAUGC GGCUCC 129 miR-24/24ab/24-3p GGCUCA UGAGCC GGAGCC AAUGCC 130 miR-33a-3p/365/365-3p AAUGCC GGCAUU GGCAUU CUACAA 131 miR-139-5p CUACAG CUGUAG UUGUAG GCUGGG 132 miR-138/138ab GCUGGU ACCAGC CCCAGC GAGAUU 133 miR-143/1721/4770 GAGAUG CAUCUC AAUCUC AUUGCC 134 miR-25/32/92abc/363/363-3p/367 AUUGCA UGCAAU GGCAAU GAGUGG 135 miR-574-5p GAGUGU ACACUC CCACUC GGAAGG 136 miR-7/7ab GGAAGA UCUUCC CCUUCC UCCAGG 137 miR-145 UCCAGU ACUGGA CCUGGA AUGGCC 138 miR-135ab/135a-5p AUGGCU AGCCAU GGCCAU CAGUGG 139 miR-148ab-3p/152 CAGUGC GCACUG CCACUG AGGAGG 140 miR-28-5p/708/1407/1653/3139 AGGAGC GCUCCU CCUCCU AGUGCC 141 miR-130ac/301ab/301b/301b-3p/454/721/4295/3666 AGUGCA UGCACU GGCACU GGGUAA 142 miR-3132 GGGUAG CUACCC UUACCC UAAUGG 143 miR-155 UAAUGC GCAUUA CCAUUA UCAUAA 144 miR-485-3p UCAUAC GUAUGA UUAUGA AACAGG 145 miR-132/212/212-3p AACAGU ACUGUU CCUGUU UAAAGG 146 hsa-miR-9-3p UAAAGC GCUUUA CCUUUA UAUAAA 147 miR-374ab UAUAAU AUUAUA UUUAUA AGCCCC 148 miR-129-3p/129ab-3p/129-1-3p/129-2-3p AGCCCU AGGGCU GGGGCU AUUAUU 149 hsa-miR-126-5p AUUAUU AAUAAU AAUAAU AUGACC 150 miR-425/425-5p/489 AUGACA UGUCAU GGUCAU GCUCGG 151 miR-423-3p GCUCGG CCGAGC CCGAGC AGCUUU 152 miR-21/590-5p AGCUUA UAAGCU AAAGCU GGCAAA 153 miR-31 GGCAAG CUUGCC UUUGCC CUGUAA 154 hsa-miR-20b-3p CUGUAG CUACAG UUACAG UAUACC 155 hsa-let-7d-3p UAUACG CGUAUA GGUAUA AACGGG 156 miR-191 AACGGA UCCGUU CCCGUU AAGGUU 157 miR-18ab/4735-3p AAGGUG CACCUU AACCUU AUAAUU 158 miR-369-3p AUAAUA UAUUAU AAUUAU GGGAUU 159 hsa-miR-5187-5p GGGAUG CAUCCC AAUCCC AAGUUU 160 miR-382 AAGUUG CAACUU AAACUU GAGGCC 161 miR-485-5p/1698/1703/1962 GAGGCU AGCCUC GGCCUC AUCAUU 162 hsa-miR-136-3p AUCAUC GAUGAU AAUGAU AGAUGG 163 miR-576-3p AGAUGU ACAUCU CCAUCU UCCCUU 164 miR-204/204b/211 UCCCUU AAGGGA AAGGGA GAGACC 165 miR-769-5p GAGACC GGUCUC GGUCUC GGGGUU 166 miR-342-5p/4664-5p GGGGUG CACCCC AACCCC UAUCAA 167 miR-361-5p UAUCAG CUGAUA UUGAUA CAGUAA 168 miR-199ab-3p/3129-5p CAGUAG CUACUG UUACUG GUAGUU 169 miR-142-3p GUAGUG CACUAC AACUAC GGUUUU 170 miR-299-5p/3563-5p GGUUUA UAAACC AAAACC ACUGGG 171 miR-193/193b/193a-3p ACUGGC GCCAGU CCCAGU AAUAUU 172 hsa-miR-1277-5p AAUAUA UAUAUU AAUAUU AGUGGG 173 miR-140/140-5p/876-3p/1244 AGUGGU ACCACU CCCACU UUUCAA 174 hsa-miR-30a/d/e-3p UUUCAG CUGAAA UUGAAA UGCGCC 175 hsa-let-7i-3p UGCGCA UGCGCA GGCGCA GGUUAA 176 miR-409-5p/409a GGUUAC GUAACC UUAACC GGUAGG 177 miR-379/1193-5p/3529 GGUAGA UCUACC CCUACC CUCCAA 178 miR-136 CUCCAU AUGGAG UUGGAG AGGUUU 179 miR-154/872 AGGUUA UAACCU AAACCU GUUGCC 180 miR-4684-3p GUUGCA UGCAAC GGCAAC CCCCCC 181 miR-361-3p CCCCCA UGGGGG GGGGGG CAAGAA 182 miR-335/335-5p CAAGAG CUCUUG UUCUUG GAGGGG 183 miR-423a/423-5p/3184/3573-5p GAGGGG CCCCUC CCCCUC CUCAAA 184 miR-371/373/371b-5p CUCAAA UUUGAG UUUGAG GAGGAA 185 miR-1185/3679-5p GAGGAU AUCCUC UUCCUC CAAAAA 186 miR-3613-3p CAAAAA UUUUUG UUUUUG AAGUGG 187 miR-93/93a/105/106a/291a- AAGUGC GCACUU CCACUU 3p/294/295/302abcde/372/373/428/519a/520be/520 acd-3p/1378/1420ac GGAUUU 188 miR-876-5p/3167 GGAUUU AAAUCC AAAUCC ACACAA 189 miR-329/329ab/362-3p ACACAC GUGUGU UUGUGU UACAGG 190 miR-582-5p UACAGU ACUGUA CCUGUA GAGAAA 191 miR-146ac/146b-5p GAGAAC GUUCUC UUUCUC AUGUAA 192 miR-380/380-3p AUGUAA UUACAU UUACAU ACAUCC 193 miR-499-3p/499a-3p ACAUCA UGAUGU GGAUGU CGACCC 194 miR-551a CGACCC GGGUCG GGGUCG AUAAAA 195 miR-142-5p AUAAAG CUUUAU UUUUAU CUGCAA 196 hsa-miR-17-3p CUGCAG CUGCAG UUGCAG CCAGUU 197 miR-199ab-5p CCAGUG CACUGG AACUGG GUGACC 198 miR-542-3p GUGACA UGUCAC GGUCAC ACGUAA 199 miR-1277 ACGUAG CUACGU UUACGU GACCGG 200 hsa-miR-29c-5p GACCGA UCGGUC CCGGUC GAUAUU 201 miR-3145-3p GAUAUU AAUAUC AAUAUC CGCACC 202 hsa-miR-106b-3p CGCACU AGUGCG GGUGCG GUUCUU 203 hsa-miR-22-5p GUUCUU AAGAAC AAGAAC GCGGGG 204 miR-744/1716 GCGGGG CCCCGC CCCCGC CCGUGG 205 hsa-miR-132-5p CCGUGG CCACGG CCACGG UGAAAA 206 miR-488 UGAAAG CUUUCA UUUUCA AUGCAA 207 miR-501-3p/502-3p/500/502a AUGCAC GUGCAU UUGCAU CCUGUU 208 miR-486-5p/3107 CCUGUA UACAGG AACAGG UUUGCC 209 miR-450a/451a UUUGCG CGCAAA GGCAAA UGGGAA 210 hsa-miR-30c-3p UGGGAG CUCCCA UUCCCA UAAGAA 211 miR-499-5p UAAGAC GUCUUA UUCUUA UCAACC 212 miR-421 UCAACA UGUUGA GGUUGA UCACCC 213 miR-197 UCACCA UGGUGA GGGUGA GGGCCC 214 miR-296-5p GGGCCC GGGCCC GGGCCC CUCUGG 215 miR-326/330/330-5p CUCUGG CCAGAG CCAGAG CAGCAA 216 miR-214/761/3619-5p CAGCAG CUGCUG UUGCUG CUGGGG 217 miR-612/1285/3187-5p CUGGGC GCCCAG CCCCAG AAUGUU 218 miR-409-3p AAUGUU AACAUU AACAUU CUGGAA 219 miR-378/422a/378bcdefhi CUGGAC GUCCAG UUCCAG CUCACC 220 miR-342-3p CUCACA UGUGAG GGUGAG ACAAUU 221 miR-338-5p ACAAUA UAUUGU AAUUGU GGGGGG 222 miR-625 GGGGGA UCCCCC CCCCCC AAUACC 223 miR-200bc/429/548a AAUACU AGUAUU GGUAUU UAGAUU 224 hsa-miR-376a-5p UAGAUU AAUCUA AAUCUA UAUGGG 225 miR-584 UAUGGU ACCAUA CCCAUA AGUAGG 226 miR-411 AGUAGA UCUACU CCUACU UGAAGG 227 miR-573/3533/3616-5p/3647-5p UGAAGU ACUUCA CCUUCA CCAUUU 228 miR-885-5p CCAUUA UAAUGG AAAUGG AAGCUU 229 hsa-miR-99-3p AAGCUC GAGCUU AAGCUU GGUGGG 230 miR-876-3p GGUGGU ACCACC CCCACC AUGUCC 231 miR-654-3p AUGUCU AGACAU GGACAU CCGUCC 232 hsa-miR-340-3p CCGUCU AGACGG GGACGG CACUUU 233 miR-3614-5p CACUUG CAAGUG AAAGUG GUGUUU 234 hsa-miR-124-5p GUGUUC GAACAC AAACAC GUGGGG 235 miR-491-5p GUGGGG CCCCAC CCCCAC UUGGCC 236 miR-96/507/1271 UUGGCA UGCCAA GGCCAA AAAACC 237 miR-548a-3p/548ef/2285a AAAACU AGUUUU GGUUUU AAUUUU 238 hsa-miR-32-3p AAUUUA UAAAUU AAAAUU AGCAAA 239 miR-3942-5p/4703-5p AGCAAU AUUGCU UUUGCU AGGCAA 240 miR-34b/449c/1360/2682 AGGCAG CUGCCU UUGCCU GGGUUU 241 hsa-miR-23a/b-5p GGGUUC GAACCC AAACCC AUCCUU 242 miR-362-5p/500b AUCCUU AAGGAU AAGGAU UCACUU 243 miR-677/4420 UCACUG CAGUGA AAGUGA AGAUAA 244 miR-577 AGAUAA UUAUCU UUAUCU GUUGUU 245 miR-3613-5p GUUGUA UACAAC AACAAC GAUCGG 246 miR-369-5p GAUCGA UCGAUC CCGAUC CUCCCC 247 miR-150/5127 CUCCCA UGGGAG GGGGAG UUCUGG 248 miR-544/544ab/544-3p UUCUGC GCAGAA CCAGAA CUGAUU 249 hsa-miR-29a-5p CUGAUU AAUCAG AAUCAG CAGGAA 250 miR-873 CAGGAA UUCCUG UUCCUG AGCCUU 251 miR-3614-3p AGCCUU AAGGCU AAGGCU AAAGAA 252 miR-186 AAAGAA UUCUUU UUCUUU CACUCC 253 miR-483-3p CACUCC GGAGUG GGAGUG UUAUCC 254 hsa-miR-374a-3p UUAUCA UGAUAA GGAUAA AGGUAA 255 miR-196abc AGGUAG CUACCU UUACCU GAUUCC 256 hsa-miR-145-3p GAUUCC GGAAUC GGAAUC UGGUUU 257 hsa-miR-29b-2-5p UGGUUU AAACCA AAACCA CCUGGG 258 hsa-miR-221-5p CCUGGC GCCAGG CCCAGG CCAAUU 259 miR-323b-3p CCAAUA UAUUGG AAUUGG GUCAUU 260 miR-616 GUCAUU AAUGAC AAUGAC CAAAGG 261 miR-330-3p CAAAGC GCUUUG CCUUUG AACAAA 262 hsa-miR-7-3p AACAAA UUUGUU UUUGUU CGUGUU 263 miR-187 CGUGUC GACACG AACACG CUAUUU 264 hsa-miR-26a-3p CUAUUC GAAUAG AAAUAG ACUGUU 265 miR-452/4676-3p ACUGUU AACAGU AACAGU UUUUUU 266 miR-129-5p/129ab-5p UUUUUG CAAAAA AAAAAA GUCAGG 267 miR-223 GUCAGU ACUGAC CCUGAC GCCAGG 268 miR-4755-3p GCCAGG CCUGGC CCUGGC CCCGUU 269 miR-1247 CCCGUC GACGGG AACGGG AACUAA 270 miR-3129-3p AACUAA UUAGUU UUAGUU UUUUCC 271 hsa-miR-335-3p UUUUCA UGAAAA GGAAAA CGGGGG 272 miR-542-5p CGGGGA UCCCCG CCCCCG CCAUCC 273 hsa-miR-181a-3p CCAUCG CGAUGG GGAUGG CCCAAA 274 hsa-miR-186-3p CCCAAA UUUGGG UUUGGG GAGCUU 275 hsa-miR-27b-5p GAGCUU AAGCUC AAGCUC UUAUGG 276 miR-491-3p UUAUGC GCAUAA CCAUAA GGCUGG 277 miR-4687-3p GGCUGU ACAGCC CCAGCC AGUUAA 278 hsa-miR-101-5p AGUUAU AUAACU UUAACU GAUCAA 279 miR-4772-5p GAUCAG CUGAUC UUGAUC UCCUAA 280 miR-337-3p UCCUAU AUAGGA UUAGGA GUGUAA 281 hsa-miR-223-5p GUGUAU AUACAC UUACAC CAAUAA 282 hsa-miR-16/195-3p CAAUAU AUAUUG UUAUUG UCGUGG 283 miR-3677-3p UCGUGG CCACGA CCACGA GGAGGG 284 hsa-miR-766-5p GGAGGA UCCUCC CCCUCC AUGUGG 285 miR-299/299-3p/3563-3p AUGUGG CCACAU CCACAU GCUUUU 286 miR-3140-3p GCUUUU AAAAGC AAAAGC AUGCCC 287 miR-532-5p/511 AUGCCU AGGCAU GGGCAU GCCUAA 288 hsa-miR-24-5p GCCUAC GUAGGC UUAGGC AUUCUU 289 miR-4778-5p AUUCUG CAGAAU AAGAAU GACACC 290 miR-642b GACACA UGUGUC GGUGUC AGACGG 291 miR-483-5p AGACGG CCGUCU CCGUCU GCACCC 292 miR-767-5p GCACCA UGGUGC GGGUGC GCUAUU 293 hsa-miR-31-3p GCUAUG CAUAGC AAUAGC ACGCUU 294 miR-574-3p ACGCUC GAGCGU AAGCGU AAGGAA 295 miR-3173-3p AAGGAG CUCCUU UUCCUU GGGAGG 296 miR-2127/4728-5p GGGAGG CCUCCC CCUCCC GCUUCC 297 hsa-miR-103a-2-5p GCUUCU AGAAGC GGAAGC AACACC 298 miR-3591-3p AACACC GGUGUU GGUGUU ACUAUU 299 hsa-miR-625-3p ACUAUA UAUAGU AAUAGU GAAUCC 300 hsa-miR-15b-3p GAAUCA UGAUUC GGAUUC AAAUGG 301 miR-522/518e/1422p AAAUGG CCAUUU CCAUUU AAAAAA 302 miR-548d-3p/548acbz AAAAAC GUUUUU UUUUUU UCAUCC 303 hsa-miR-452-3p UCAUCU AGAUGA GGAUGA UGACCC 304 miR-192/215 UGACCU AGGUCA GGGUCA UAGCAA 305 miR-1551/4524 UAGCAG CUGCUA UUGCUA UCGGGG 306 hsa-miR-425-3p UCGGGA UCCCGA CCCCGA AUCUGG 307 miR-3126-3p AUCUGG CCAGAU CCAGAU CACAAA 308 hsa-miR-125b-2-3p CACAAG CUUGUG UUUGUG CUGCCC 309 miR-324-3p/1913 CUGCCC GGGCAG GGGCAG AUCUUU 310 hsa-miR-141-5p AUCUUC GAAGAU AAAGAU GGGACC 311 hsa-miR-365a/b-5p GGGACU AGUCCC GGUCCC CUGGUU 312 hsa-miR-29b-1-5p CUGGUU AACCAG AACCAG GGUUGG 313 miR-563/380-5p GGUUGA UCAACC CCAACC UUGAGG 314 miR-1304 UUGAGG CCUCAA CCUCAA UCUCUU 315 miR-216c/1461/4684-5p UCUCUA UAGAGA AAGAGA UUUUAA 316 hsa-miR-2681-5p UUUUAC GUAAAA UUAAAA GUAACC 317 miR-194 GUAACA UGUUAC GGUUAC AGGGUU 318 miR-296-3p AGGGUU AACCCU AACCCU AUUUCC 319 hsa-miR-205-3p AUUUCA UGAAAU GGAAAU ACUCAA 320 miR-888 ACUCAA UUGAGU UUGAGU ACAUGG 321 miR-4802-3p ACAUGG CCAUGU CCAUGU UGUACC 322 hsa-let-7a/g-3p UGUACA UGUACA GGUACA GGGCUU 323 miR-762/4492/4498 GGGCUG CAGCCC AAGCCC UGUUGG 324 hsa-miR-744-3p UGUUGC GCAACA CCAACA AGUUCC 325 hsa-miR-148b-5p AGUUCU AGAACU GGAACU UUGACC 326 miR-514/514b-3p UUGACA UGUCAA GGUCAA ACUAGG 327 miR-28-3p ACUAGA UCUAGU CCUAGU GUGCCC 328 miR-550a GUGCCU AGGCAC GGGCAC CGGGUU 329 hsa-miR-125b-1-3p CGGGUU AACCCG AACCCG AUUCAA 330 hsa-miR-506-5p AUUCAG CUGAAU UUGAAU CACCUU 331 hsa-miR-1306-5p CACCUC GAGGUG AAGGUG CCUUGG 332 miR-3189-3p CCUUGG CCAAGG CCAAGG GGUGCC 333 miR-675-5p/4466 GGUGCG CGCACC GGCACC AAUCAA 334 hsa-miR-34a-3p AAUCAG CUGAUU UUGAUU CCCUAA 335 hsa-miR-454-5p CCCUAU AUAGGG UUAGGG ACUGCC 336 miR-509-5p/509-3-5p/4418 ACUGCA UGCAGU GGCAGU GUUUUU 337 hsa-miR-19a/b-5p GUUUUG CAAAAC AAAAAC UUCCCC 338 miR-4755-5p UUCCCU AGGGAA GGGGAA CUGCUU 339 hsa-miR-93-3p CUGCUG CAGCAG AAGCAG ACCCAA 340 miR-3130-5p/4482 ACCCAG CUGGGU UUGGGU CCAGAA 341 hsa-miR-488-5p CCAGAU AUCUGG UUCUGG UCCUGG 342 hsa-miR-378a-5p UCCUGA UCAGGA CCAGGA AGCCAA 343 miR-575/4676-5p AGCCAG CUGGCU UUGGCU CUCGGG 344 miR-1307 CUCGGC GCCGAG CCCGAG UUCAGG 345 miR-3942-3p UUCAGA UCUGAA CCUGAA UGUUCC 346 miR-4677-5p UGUUCU AGAACA GGAACA GAGCGG 347 miR-339-3p GAGCGC GCGCUC CCGCUC AAGAAA 348 miR-548b-3p AAGAAC GUUCUU UUUCUU GUUCCC 349 hsa-miR-642b-5p GUUCCC GGGAAC GGGAAC AUCCCC 350 miR-188-5p AUCCCU AGGGAU GGGGAU AACCCC 351 hsa-miR-652-5p AACCCU AGGGUU GGGGUU AGUCCC 352 miR-2114 AGUCCC GGGACU GGGACU GUGGCC 353 miR-3688-5p GUGGCA UGCCAC GGCCAC AGGCCC 354 hsa-miR-15a-3p AGGCCA UGGCCU GGGCCU ACCAUU 355 hsa-miR-181c-3p ACCAUC GAUGGU AAUGGU GGAGUU 356 miR-122/122a/1352 GGAGUG CACUCC AACUCC UAUUAA 357 miR-556-3p UAUUAC GUAAUA UUAAUA AUGGUU 358 hsa-miR-218-2-3p AUGGUU AACCAU AACCAU CUUGUU 359 miR-643 CUUGUA UACAAG AACAAG ACCACC 360 miR-140-3p ACCACA UGUGGU GGUGGU AGUGAA 361 miR-1245 AGUGAU AUCACU UUCACU AUCAGG 362 hsa-miR-2115-3p AUCAGA UCUGAU CCUGAU AAAGCC 363 miR-518bcf/518a-3p/518d-3p AAAGCG CGCUUU GGCUUU ACCUUU 364 miR-3200-3p ACCUUG CAAGGU AAAGGU CAACAA 365 miR-545/3065/3065-5p CAACAA UUGUUG UUGUUG CUUCUU 366 miR-1903/4778-3p CUUCUU AAGAAG AAGAAG CUUAAA 367 hsa-miR-302a-5p CUUAAA UUUAAG UUUAAG UGAAUU 368 hsa-miR-183-3p UGAAUU AAUUCA AAUUCA GGGGAA 369 miR-3144-5p GGGGAC GUCCCC UUCCCC AACUGG 370 miR-582-3p AACUGG CCAGUU CCAGUU AAGAUU 371 miR-4662a-3p AAGAUA UAUCUU AAUCUU CCUGAA 372 miR-3140-5p CCUGAA UUCAGG UUCAGG UGCAAA 373 hsa-miR-106a-3p UGCAAU AUUGCA UUUGCA AUAGGG 374 hsa-miR-135a-3p AUAGGG CCCUAU CCCUAU CUGACC 375 miR-345/345-5p CUGACU AGUCAG GGUCAG CAGGUU 376 miR-125a-3p/1554 CAGGUG CACCUG AACCUG ACUCCC 377 miR-3145-5p ACUCCA UGGAGU GGGAGU UGUCCC 378 miR-676 UGUCCU AGGACA GGGACA GCCCUU 379 miR-3173-5p GCCCUG CAGGGC AAGGGC AGAGUU 380 hsa-miR-5586-3p AGAGUG CACUCU AACUCU CCGAGG 381 miR-615-3p CCGAGC GCUCGG CCUCGG AUGGAA 382 miR-3688-3p AUGGAA UUCCAU UUCCAU UAGCCC 383 miR-4662a-5p UAGCCA UGGCUA GGGCUA UGCCAA 384 miR-4659ab-5p UGCCAU AUGGCA UUGGCA AUCCAA 385 hsa-miR-5586-5p AUCCAG CUGGAU UUGGAU ACUCUU 386 hsa-miR-514a-5p ACUCUG CAGAGU AAGAGU ACCCUU 387 miR-10abc/10a-5p ACCCUG CAGGGU AAGGGU ACUGAA 388 hsa-miR-888-3p ACUGAC GUCAGU UUCAGU UCAGGG 389 miR-3127-5p UCAGGG CCCUGA CCCUGA GAUUGG 390 miR-508-3p GAUUGU ACAAUC CCAAUC GGGGCC 391 hsa-miR-185-3p GGGGCU AGCCCC GGCCCC GUCUUU 392 hsa-miR-200c-5p,hsa-miR-550a-3p GUCUUA UAAGAC AAAGAC UCUCAA 393 miR-513c/514b-5p UCUCAA UUGAGA UUGAGA AACCUU 394 miR-490-3p AACCUG CAGGUU AAGGUU CUGAAA 395 hsa-miR-5187-3p CUGAAU AUUCAG UUUCAG CUCAGG 396 miR-3664-3p CUCAGG CCUGAG CCUGAG GCCCCC 397 miR-3189-5p GCCCCA UGGGGC GGGGGC GAAGUU 398 miR-4670-3p GAAGUU AACUUC AACUUC CAAAUU 399 miR-105/105ab CAAAUG CAUUUG AAUUUG UGUAGG 400 hsa-miR-135b-3p UGUAGG CCUACA CCUACA UUUGUU 401 hsa-miR-5010-3p UUUGUG CACAAA AACAAA GAAGGG 402 miR-493/493b GAAGGU ACCUUC CCCUUC CUCCGG 403 miR-3605-3p CUCCGU ACGGAG CCGGAG UCCCAA 404 miR-188-3p UCCCAC GUGGGA UUGGGA UGCUAA 405 hsa-miR-449c-3p UGCUAG CUAGCA UUAGCA CAAGGG 406 miR-4761-5p CAAGGU ACCUUG CCCUUG AAGUCC 407 miR-224 AAGUCA UGACUU GGACUU GUCUAA 408 miR-4796-5p GUCUAU AUAGAC UUAGAC AAAUCC 409 hsa-miR-551b-5p AAAUCA UGAUUU GGAUUU AUGAGG 410 miR-556-5p AUGAGC GCUCAU CCUCAU ACGCCC 411 hsa-miR-122-3p ACGCCA UGGCGU GGGCGU CUGUGG 412 miR-4677-3p CUGUGA UCACAG CCACAG UAGAGG 413 miR-877 UAGAGG CCUCUA CCUCUA UUCUAA 414 miR-576-5p UUCUAA UUAGAA UUAGAA CAUGGG 415 miR-490-5p CAUGGA UCCAUG CCCAUG CAGAAA 416 hsa-miR-589-3p CAGAAC GUUCUG UUUCUG GAAGCC 417 miR-4786-3p GAAGCC GGCUUC GGCUUC UUAGCC 418 hsa-miR-374b-3p UUAGCA UGCUAA GGCUAA CUGUUU 419 hsa-miR-26b-3p CUGUUC GAACAG AAACAG AGGGCC 420 miR-3158-3p AGGGCU AGCCCU GGCCCU UAGGCC 421 miR-4423-3p UAGGCA UGCCUA GGCCUA UCUAGG 422 miR-518d-5p/519bc-5p520c-5p/523b/526a UCUAGA UCUAGA CCUAGA GCCCGG 423 miR-4707-3p GCCCGC GCGGGC CCGGGC AAAUUU 424 hsa-miR-10a-3p AAAUUC GAAUUU AAAUUU UCUUGG 425 miR-526b UCUUGA UCAAGA CCAAGA CUUCAA 426 hsa-miR-676-5p CUUCAA UUGAAG UUGAAG CCUCCC 427 hsa-miR-660-3p CCUCCU AGGAGG GGGAGG UUGGAA 428 hsa-miR-5004-3p UUGGAU AUCCAA UUCCAA GGGUCC 429 miR-193a-5p GGGUCU AGACCC GGACCC UCAGUU 430 hsa-miR-222-5p UCAGUA UACUGA AACUGA AGGAUU 431 miR-4661-3p AGGAUC GAUCCU AAUCCU GGCGGG 432 hsa-miR-25-5p GGCGGA UCCGCC CCCGCC AGCGAA 433 miR-4670-5p AGCGAC GUCGCU UUCGCU UUGGUU 434 miR-659 UUGGUU AACCAA AACCAA GCUCUU 435 miR-1745/3194-3p GCUCUG CAGAGC AAGAGC GGUUCC 436 hsa-miR-182-3p GGUUCU AGAACC GGAACC GCAGAA 437 miR-298/2347/2467-3p GCAGAG CUCUGC UUCUGC CUCUUU 438 hsa-miR-130b-5p CUCUUU AAAGAG AAAGAG GCGGUU 439 miR-4746-3p GCGGUG CACCGC AACCGC GCGCGG 440 miR-1893/2277-5p GCGCGG CCGCGC CCGCGC GGACCC 441 miR-3619-3p GGACCA UGGUCC GGGUCC CUACUU 442 hsa-miR-138-1-3p CUACUU AAGUAG AAGUAG AUGCUU 443 miR-4728-3p AUGCUG CAGCAU AAGCAU CCCCUU 444 miR-3127-3p CCCCUU AAGGGG AAGGGG CCGGUU 445 miR-671-3p CCGGUU AACCGG AACCGG CAGGGG 446 hsa-miR-211-3p CAGGGA UCCCUG CCCCUG GAGCCC 447 hsa-miR-2114-3p GAGCCU AGGCUC GGGCUC CCUCUU 448 hsa-miR-877-3p CCUCUU AAGAGG AAGAGG UCAGCC 449 miR-3157-5p UCAGCC GGCUGA GGCUGA UCCUUU 450 miR-502-5p UCCUUG CAAGGA AAAGGA AAUCCC 451 miR-500a AAUCCU AGGAUU GGGAUU AAACUU 452 m iR-548g AAACUG CAGUUU AAGUUU AACGCC 453 miR-523 AACGCG CGCGUU GGCGUU CAGUUU 454 hsa-miR-584-3p CAGUUC GAACUG AAACUG CCUUCC 455 miR-205/205ab CCUUCA UGAAGG GGAAGG CAUCCC 456 miR-4793-5p CAUCCU AGGAUG GGGAUG GGGUGG 457 hsa-miR-363-5p GGGUGG CCACCC CCACCC GCCUGG 458 hsa-miR-214-5p GCCUGU ACAGGC CCAGGC UUCCAA 459 miR-3180-5p UUCCAG CUGGAA UUGGAA UGGGGG 460 miR-1404/2110 UGGGGA UCCCCA CCCCCA UGCCCC 461 miR-3157-3p UGCCCU AGGGCA GGGGCA CUGCGG 462 hsa-miR-191-3p CUGCGC GCGCAG CCGCAG UGGGUU 463 miR-1346/3940-5p/4507 UGGGUU AACCCA AACCCA CGGUCC 464 miR-4746-5p CGGUCC GGACCG GGACCG ACGCGG 465 miR-3939 ACGCGC GCGCGU CCGCGU CCACUU 466 hsa-miR-181a-2-3p CCACUG CAGUGG AAGUGG UGCACC 467 hsa-miR-500a-3p UGCACC GGUGCA GGUGCA CGACAA 468 hsa-miR-196b-3p CGACAG CUGUCG UUGUCG UGUAUU 469 hsa-miR-675-3p UGUAUG CAUACA AAUACA GCAAAA 470 hsa-miR-548aj/g/x-5p GCAAAA UUUUGC UUUUGC UUCUUU 471 miR-4659ab-3p UUCUUC GAAGAA AAAGAA UCUGCC 472 hsa-miR-5001-3p UCUGCC GGCAGA GGCAGA CCCGGG 473 hsa-miR-1247-3p CCCGGG CCCGGG CCCGGG CCCCGG 474 miR-2890/4707-5p CCCCGG CCGGGG CCGGGG UGGUAA 475 hsa-miR-150-3p UGGUAC GUACCA UUACCA UUCUCC 476 hsa-miR-629-3p UUCUCC GGAGAA GGAGAA GACAGG 477 miR-2277-3p GACAGC GCUGUC CCUGUC GAGCAA 478 miR-3547/3663-3p GAGCAC GUGCUC UUGCUC AUCACC 479 miR-34bc-3p AUCACU AGUGAU GGUGAU AAGCGG 480 miR-518ef AAGCGC GCGCUU CCGCUU UGGCCC 481 miR-3187-3p UGGCCA UGGCCA GGGCCA CGUUGG 482 miR-1306/1306-3p CGUUGG CCAACG CCAACG GCACGG 483 miR-3177-3p GCACGG CCGUGC CCGUGC GGAAUU 484 miR-1ab/206/613 GGAAUG CAUUCC AAUUCC CACAGG 485 miR-128/128ab CACAGU ACUGUG CCUGUG UAGGGG 486 miR-1296 UAGGGC GCCCUA CCCCUA ACGUCC 487 miR-598/598-3p ACGUCA UGACGU GGACGU UGAACC 488 miR-887 UGAACG CGUUCA GGUUCA CAUACC 489 miR-1-5p CAUACU AGUAUG GGUAUG ACAUAA 490 miR-376c/741-5p ACAUAG CUAUGU UUAUGU UAAUAA 491 miR-374c/655 UAAUAC GUAUUA UUAUUA GAAACC 492 miR-494 GAAACA UGUUUC GGUUUC UUAGGG 493 miR-651 UUAGGA UCCUAA CCCUAA UGCAGG 494 miR-1301/5047 UGCAGC GCUGCA CCUGCA GCGAGG 495 miR-381-5p GCGAGG CCUCGC CCUCGC AAUCUU 496 miR-216a AAUCUC GAGAUU AAGAUU AUACAA 497 miR-300/381/539-3p AUACAA UUGUAU UUGUAU CGCCCC 498 miR-1249 CGCCCU AGGGCG GGGGCG UCAUUU 499 miR-579 UCAUUU AAAUGA AAAUGA AUAUUU 500 miR-656 AUAUUA UAAUAU AAAUAU UCAUGG 501 miR-433 UCAUGA UCAUGA CCAUGA UUCCGG 502 miR-1180 UUCCGG CCGGAA CCGGAA GUGUCC 503 miR-597/1970 GUGUCA UGACAC GGACAC UAUAUU 504 miR-190a-3p UAUAUA UAUAUA AAUAUA AAACCC 505 miR-1537 AAACCG CGGUUU GGGUUU GGCCCC 506 miR-874-5p GGCCCC GGGGCC GGGGCC AUAUAA 507 miR-410/344de/344b-1-3p AUAUAA UUAUAU UUAUAU CCUGCC 508 miR-370 CCUGCU AGCAGG GGCAGG GAAUUU 509 miR-219-2-3p/219-3p GAAUUG CAAUUC AAAUUC CACCCC 510 miR-3620 CACCCU AGGGUG GGGGUG GACCCC 511 miR-504/4725-5p GACCCU AGGGUC GGGGUC GAUGUU 512 miR-2964/2964a-5p GAUGUC GACAUC AACAUC UUGGGG 513 miR-450a-2-3p UUGGGG CCCCAA CCCCAA UGUCUU 514 miR-511 UGUCUU AAGACA AAGACA GACUUU 515 miR-6505-3p GACUUC GAAGUC AAAGUC ACGGUU 516 miR-433-5p ACGGUG CACCGU AACCGU CGGCUU 517 miR-6741-3p CGGCUC GAGCCG AAGCCG AGGUCC 518 miR-370-5p AGGUCA UGACCU GGACCU CGCGGG 519 miR-579-5p CGCGGU ACCGCG CCCGCG GUGGAA 520 miR-376c-5p,miR-376b-5p GUGGAU AUCCAC UUCCAC ACAGGG 521 miR-552/3097-5p ACAGGU ACCUGU CCCUGU CAGUCC 522 miR-1910 CAGUCC GGACUG GGACUG UUGUGG 523 miR-758 UUGUGA UCACAA CCACAA GGCCUU 524 miR-6735-3p GGCCUG CAGGCC AAGGCC GUAGAA 525 miR-376a-2-5p GUAGAU AUCUAC UUCUAC GGGCGG 526 miR-585 GGGCGU ACGCCC CCGCCC AACCGG 527 miR-451 AACCGU ACGGUU CCGGUU UAUUGG 528 miR-137/137ab UAUUGC GCAAUA CCAAUA
[0368] Through the Ago HITS-CLIP assay in the above example, miRNA binding to a non-canonical bulge target in several tissue cells was identified, and when the present invention for modifying miRNA to recognize a non-canonical nucleation bulge site as a canonical seed site is applied, a total of 426 sequences (BS sequences) were prepared, and therefore, technology for exhibiting only the function of suppressing the non-canonical target of miRNA was completed.
EXAMPLE 25
Analysis of Sequence Variation in miRNA Sequence Database (TCGA) of Cancer Patients, and Thereby Identifying miRNA Binding to Non-Canonical G:A Wobble Seed Site
[0369] According to the above examples, it was confirmed that miRNA binds to a non-canonically G:A wobble site, thereby inhibiting the expression of a target gene, and therefore, in the present invention, a miRNA sequence modification method (miRNA-BS) of modifying a sequence to recognize a non-canonical G:A wobble site as a canonical seed site was developed. If a miRNA function is changed through wobble pairing between G in the conventional miRNA seed sequence and A of target mRNA, based on the idea that such a mechanism can be shown in a disease such as a tumor through sequence variation of miRNA, a result of sequencing miRNA from tissue of a cancer patient was analyzed (miRNA-Seq). Here, as 8,105 sequences were obtained from the miRNA sequencing-related TCGA database of cancer patients, and additionally, 468 sequences were obtained through cancer-related miRNA sequencing from the Gene Expression Omnibus (GEO) database, a total of 8,573 tumor miRNA sequencing results obtained thereby were mapped using the bowtie2 program in the same manner as in Example 10, and then sites with variations were analyzed. At this time, the miRNA sequencing data used in the analysis for each cancer type is as shown in
[0370] As a result of examining the distribution of the most frequent variations (top 10,000) and the least variations (bottom 10,000) by classifying variation fractions by position based on the 5′ end of miRNA (
[0371] This may be a phenomenon in which G of miRNA recognizing the non-canonical G:A wobble noted by the inventors is changed to U to more tightly bind to A of the target mRNA on the opposite side, and as a result of analyzing which bases Gs, which show the higher variation rate, were changed to (
TABLE-US-00004 TABLE 4 Sequence SEQ G > T (G > U) ID NOs miRNA name Seed Position read/patient UGAAUG 529 hsa-miR-1-3p GGAAUG 2 17.1 UUAACA 530 hsa-miR-194-5p GUAACA 2 96.3 UGGUCU 531 hsa-miR-193a-5p GGGUCU 2 12 UAAUCA 532 hsa-miR-15b-3p GAAUCA 2 2.2 UUCUUA 533 hsa-miR-200c-5p GUCUUA 2 4.2 UCCUGU 534 hsa-miR-214-5p GCCUGU 2 2.8 UUGACU 535 hsa-miR-134-5p GUGACU 2 10.3 UAUUCC 536 hsa-miR-145-3p GAUUCC 2 2.2 UUUCUU 537 hsa-miR-22-5p GUUCUU 2 2.5 UCUCGG 538 hsa-miR-423-3p GCUCGG 2 6.3 UAGACU 539 hsa-miR-873-3p GAGACU 2 4.5 UGAGUG 540 hsa-miR-122-5p GGAGUG 2 837.8 UAGAUG 541 hsa-miR-143-3p GAGAUG 2 322.7 UAGGCU 542 hsa-miR-485-5p GAGGCU 2 2.2 UGUUAC 543 hsa-miR-409-5p GGUUAC 2 8.1 UGCUCA 544 hsa-miR-24-3p GGCUCA 2 65.6 UUCAGU 545 hsa-miR-223-3p GUCAGU 2 12.7 UAUAUC 546 hsa-miR-144-5p GAUAUC 2 16.4 UGUAGA 547 hsa-miR-379-5p GGUAGA 2 47.2 UAGAAC 548 hsa-miR-146b-5p/hsa-miR-146a-5p GAGAAC 2 21.2 UAGAAA 549 hsa-miR-539-5p GAGAAA 2 3.3 UGGCCC 550 hsa-miR-296-5p GGGCCC 2 2.7 UCACCA 551 hsa-miR-767-5p GCACCA 2 10.1 UGCAGU 552 hsa-miR-34a-5p/hsa-miR-34c-5p GGCAGU 2 8.9 UAGGUA 553 hsa-let-7f-5p/hsa-let-7d-5p/hsa-let-7b-5p/hsa- GAGGUA 2 320.1 let-7a-5p/hsa-let-7e-5p/hsa-miR-202-3p/hsa- let-71-5p/hsa-miR-98-5p/hsa-let-7c-5p/hsa-let- 7g-5p UUGCCU 554 hsa-miR-1271-3p GUGCCU 2 2 UCUGGU 555 hsa-miR-138-5p GCUGGU 2 5.9 UUGCAA 556 hsa-miR-19b-3p/hsa-miR-19a-3p GUGCAA 2 4.1 UGGCUU 557 hsa-miR-27a-5p GGGCUU 2 2.2 UCCCUG 558 hsa-miR-146b-3p GCCCUG 2 7.6 UGAAGA 559 hsa-miR-7-5p GGAAGA 2 2.3 UAGGGG 560 hsa-miR-423-5p GAGGGG 2 3.7 UCAUCC 561 hsa-miR-324-5p GCAUCC 2 2.6 UGGUUU 562 hsa-miR-629-5p GGGUUU 2 3.3 UGAGAC 563 hsa-miR-139-3p GGAGAC 2 2.3 UUAAAC 564 hsa-miR-30d-5p/hsa-miR-30e-5p/hsa-miR- GUAAAC 2 1004.2 30a-5p/hsa-miR-30c-5p/hsa-miR-30b-5p UCUACA 565 hsa-miR-221-3p/hsa-miR-222-3p GCUACA 2 6.3 UAUUGG 566 hsa-miR-509-3p GAUUGG 2 54 UAGACC 567 hsa-miR-769-5p GAGACC 2 3.9 UUAGUG 568 hsa-miR-142-3p GUAGUG 2 5.2 UGAGAG 569 hsa-miR-185-5p GGAGAG 2 8.9 UAUUGU 570 hsa-miR-508-3p/hsa-miR-219a-5p GAUUGU 2 211.9 UGCAAG 571 hsa-miR-31-5p GGCAAG 2 4.2 UCAGCA 572 hsa-miR-103a-3p/hsa-miR-107 GCAGCA 2 757.3 UUGACA 573 hsa-miR-542-3p GUGACA 2 7.3 UAAUUG 574 hsa-miR-219a-2-3p GAAUUG 2 29.7 AUCACC 575 hsa-miR-29c-3p/hsa-miR-29a-3p/hsa-miR-29b-3p AGCACC 3 11 CUGGUU 576 hsa-miR-125b-1-3p CGGGUU 3 8.3 GUAAGA 577 hsa-miR-7-5p GGAAGA 3 2.3 AUUAGA 578 hsa-miR-411-5p AGUAGA 3 2.7 AUGUAG 579 hsa-miR-196a-5p/hsa-miR-196b-5p AGGUAG 3 3.3 AUGCAC 580 hsa-miR-3622a-5p AGGCAC 3 2 UUAAGC 581 hsa-miR-127-5p UGAAGC 3 4.6 AUCUGC 582 hsa-miR-22-3p AGCUGC 3 335.7 UUCAUA 583 hsa-miR-153-3p UGCAUA 3 3.1 GUGUCU 584 hsa-miR-193a-5p GGGUCU 3 5.8 GUAGAG 585 hsa-miR-185-5p GGAGAG 3 3.1 GUAAUG 586 hsa-miR-1-3p GGAAUG 3 10.2 AUCAGC 587 hsa-miR-15b-5p/hsa-miR-16-5p/hsa-miR-424-5p AGCAGC 3 5.6 UUUACA 588 hsa-let-7g-3p/hsa-miR-493-5p/hsa-let-7c-3p UGUACA 3 2.7 UUCGCA 589 hsa-let-7i-3p UGCGCA 3 2.4 UUUGCU 590 hsa-miR-218-5p UGUGCU 3 3.6 CUACCG 591 hsa-miR-1307-5p CGACCG 3 3.3 CUGAUC 592 hsa-miR-127-3p CGGAUC 3 10.5 UUUGCG 593 hsa-miR-210-3p UGUGCG 3 13.6 CUUGUC 594 hsa-miR-187-3p CGUGUC 3 2.1 UUCCAA 595 hsa-miR-192-3p UGCCAA 3 2.2 UUACCU 596 hsa-miR-192-5p UGACCU 3 73.2 GUCAGU 597 hsa-miR-34a-5p/hsa-miR-34c-5p GGCAGU 3 3.1 AUCUUA 598 hsa-miR-21-5p AGCUUA 3 738.6 UUCACC 599 hsa-miR-500a-3p UGCACC 3 2.7 GUGUUU 600 hsa-miR-629-5p GGGUUU 3 2.9 GUUAGA 601 hsa-miR-379-5p GGUAGA 3 22.1 UUAAAU 602 hsa-miR-203a-3p UGAAAU 3 200.1 GUCUCA 603 hsa-miR-24-3p GGCUCA 3 10.5 UUGGAG 604 hsa-miR-30c-2-3p UGGGAG 3 6.2 UUAAAG 605 hsa-miR-488-3p UGAAAG 3 2 AUUGCA 606 hsa-miR-301a-3p/hsa-miR-301b-3p AGUGCA 3 5 CUUACC 607 hsa-miR-126-3p CGUACC 3 15.8 GUAGUG 608 hsa-miR-122-5p GGAGUG 3 60.4 GUGCUU 609 hsa-miR-27a-5p GGGCUU 3 2.8 CUUUUC 610 hsa-miR-26b-3p CUGUUC 4 3 CUUCCC 611 hsa-miR-324-3p CUGCCC 4 2.3 CAUCAC 612 hsa-miR-3065-3p CAGCAC 4 4 GAUGGG 613 hsa-miR-423-5p GAGGGG 4 11.4 GUUUUC 614 hsa-miR-124-5p GUGUUC 4 3.3 CUUACU 615 hsa-miR-345-5p CUGACU 4 6 CCUAGC 616 hsa-miR-615-3p CCGAGC 4 2 AUUGCU 617 hsa-miR-889-5p/hsa-miR-135a-5p/hsa-miR- AUGGCU 4 15.8 135b-5p GGUUCU 618 hsa-miR-193a-5p GGGUCU 4 35.4 AAUGUG 619 hsa-miR-18a-5p AAGGUG 4 3.3 CGUGUU 620 hsa-miR-125b-1-3p CGGGUU 4 57 AGUAGC 621 hsa-miR-708-5p/hsa-miR-28-5p AGGAGC 4 10.5 AAUUCA 622 hsa-miR-224-5p AAGUCA 4 2.9 AAUCUU 623 hsa-miR-100-3p AAGCUU 4 9.5 CAUGAA 624 hsa-miR-873-5p CAGGAA 4 6.7 UAUCCA 625 hsa-miR-4662a-5p UAGCCA 4 7.5 AAUCUC 626 hsa-miR-99b-3p/hsa-miR-99a-3p AAGCUC 4 9.3 ACUGUG 627 hsa-miR-433-5p ACGGUG 4 2.3 GUUACA 628 hsa-miR-542-3p GUGACA 4 20.9 GAUGAU 629 hsa-miR-3605-5p GAGGAU 4 11.8 GCUGGG 630 hsa-miR-744-5p GCGGGG 4 4.2 UAUGGC 631 hsa-miR-1296-5p UAGGGC 4 26.2 UUUGUC 632 hsa-miR-133a-3p UUGGUC 4 11.7 AAUUUG 633 hsa-miR-382-5p AAGUUG 4 6.8 AUUACA 634 hsa-miR-425-5p AUGACA 4 20.9 GAUGUU 635 hsa-miR-377-5p GAGGUU 4 2.2 CGUAUC 636 hsa-miR-127-3p CGGAUC 4 219.1 GGUGCG 637 hsa-miR-3180-3p GGGGCG 4 2.4 GAUAUG 638 hsa-miR-143-3p GAGAUG 4 936.3 UUUUGA 639 hsa-miR-758-3p UUGUGA 4 4.9 CUUCUG 640 hsa-miR-93-3p CUGCUG 4 2.9 GGUGCC 641 hsa-miR-128-2-5p GGGGCC 4 4.2 GUUACU 642 hsa-miR-134-5p GUGACU 4 29.2 AGUUUA 643 hsa-miR-154-5p AGGUUA 4 3.5 AGUCAC 644 hsa-miR-3622a-5p AGGCAC 4 5.1 AAUGCA 645 hsa-miR-124-3p AAGGCA 4 38.7 GGUCUU 646 hsa-miR-27a-5p GGGCUU 4 5 CAUUGG 647 hsa-miR-194-3p CAGUGG 4 8.7 UUUUUC 648 hsa-miR-375 UUGUUC 4 1084.1 AAUUUC 649 hsa-miR-148a-5p AAGUUC 4 4.4 GCUCGG 650 hsa-miR-2277-5p GCGCGG 4 2.9 GAUACC 651 hsa-miR-769-5p GAGACC 4 26.3 CUUCAG 652 hsa-miR-17-3p CUGCAG 4 52.2 GAUACU 653 hsa-miR-873-3p GAGACU 4 9.9 CUUCAA 654 hsa-miR-4772-3p CUGCAA 4 2.3 AGUUUU 655 hsa-miR-329-5p AGGUUU 4 2.1 UUUGCA 656 hsa-miR-182-5p/hsa-miR-96-5p UUGGCA 4 598.4 GAUGCU 657 hsa-miR-2467-5p/hsa-miR-485-5p GAGGCU 4 12.7 CUUGCU 658 hsa-miR-149-5p CUGGCU 4 11.8 UGUUUU 659 hsa-miR-29b-2-5p UGGUUU 4 4.8 ACUCCA 660 hsa-miR-122-3p ACGCCA 4 5 AAUUGC 661 hsa-miR-302a-3p/hsa-miR-520a-3p/hsa-miR- AAGUGC 4 29.3 519b-3p/hsa-miR-520b/hsa-miR-519c-3p/hsa- miR-520c-3p/hsa-miR-519a-3p AUUCCU 662 hsa-miR-532-5p AUGCCU 4 194.9 CCUUGG 663 hsa-miR-132-5p CCGUGG 4 2.3 AAUGAU 664 hsa-miR-541-5p AAGGAU 4 2.8 CCUGUU 665 hsa-miR-671-3p CCGGUU 4 3.6 GGUCCC 666 hsa-miR-296-5p GGGCCC 4 3 AAUCGC 667 hsa-miR-518e-3p AAGCGC 4 10.8 UGUUUA 668 hsa-miR-487a-5p UGGUUA 4 2.5 GAUAAC 669 hsa-miR-589-5p/hsa-miR-146b-5p/hsa-miR- GAGAAC 4 60.3 146a-5p AGUUAG 670 hsa-miR-196b-5p/hsa-miR-196a-5p AGGUAG 4 34.4 GGUGCA 671 hsa-miR-486-3p GGGGCA 4 2 GGUUUU 672 hsa-miR-629-5p GGGUUU 4 8.7 CUUGAC 673 hsa-miR-378a-3p CUGGAC 4 105.2 GAUCUU 674 hsa-miR-27b-5p GAGCUU 4 3.2 GCUCCU 675 hsa-miR-6720-3p GCGCCU 4 2.3 ACUCUC 676 hsa-miR-574-3p ACGCUC 4 16.6 CUUAUU 677 hsa-miR-29a-5p CUGAUU 4 2.6 UGUGAG 678 hsa-miR-30c-2-3p/hsa-miR-30c-1-3p UGGGAG 4 22.2 CAUUAG 679 hsa-miR-199b-3p CAGUAG 4 32.1 GAUUGU 680 hsa-miR-574-5p GAGUGU 4 3.4 GAUAAA 681 hsa-miR-539-5p GAGAAA 4 4.7 CUUUGA 682 hsa-miR-4677-3p CUGUGA 4 3.1 AUUUCU 683 hsa-miR-654-3p AUGUCU 4 2.9 AUUGCG 684 hsa-miR-652-3p AUGGCG 4 7.8 GUUCAA 685 hsa-miR-19a-3p/hsa-miR-19b-3p GUGCAA 4 16.9 GAUGUA 686 hsa-let-7c-5p/hsa-miR-98-5p/hsa-let-7g- GAGGUA 4 1300.2 5p/hsa-let-7f-5p/hsa-miR-202-3p/hsa-let-7b- 5p/hsa-let-7e-5p/hsa-let-7a-5p/hsa-let-7d- 5p/hsa-let-7i-5p GAUCAC 687 hsa-miR-3663-3p GAGCAC 4 11.7 CAUUGC 688 hsa-miR-152-3p/hsa-miR-148b-3p/hsa-miR- CAGUGC 4 1529.4 148a-3p GGUGUU 689 hsa-miR-193b-5p GGGGUU 4 2.3 AUUCAC 690 hsa-miR-502-3p/hsa-miR-501-3p AUGCAC 4 5.5 AUUUGG 691 hsa-miR-299-3p AUGUGG 4 4.5 AGUUGU 692 hsa-miR-140-5p AGUGGU 5 4.4 UUGUCA 693 hsa-miR-96-5p/hsa-miR-182-5p UUGGCA 5 25.1 ACUUGC 694 hsa-miR-193b-3p ACUGGC 5 6.9 AAUUCC 695 hsa-miR-365a-3p AAUGCC 5 4.9 CCUUUA 696 hsa-miR-486-5p CCUGUA 5 7.4 CGGUUU 697 hsa-miR-125b-1-3p CGGGUU 5 3.9 UGUUCG 698 hsa-miR-210-3p UGUGCG 5 11.7 GAAUGU 699 hsa-miR-493-3p GAAGGU 5 2.8 AAAUUA 700 hsa-miR-548am-5p AAAGUA 5 3 UGUUCU 701 hsa-miR-218-5p UGUGCU 5 4.4 AAAUUG 702 hsa-miR-20b-5p/hsa-miR-20a-5p/hsa-miR-93- AAAGUG 5 24.7 5p/hsa-miR-17-5p/hsa-miR-106b-5p GGUUGG 703 hsa-miR-541-3p GGUGGG 5 3.8 ACUUUU 704 hsa-miR-452-5p ACUGUU 5 2.4 CCUUGC 705 hsa-miR-221-5p CCUGGC 5 6.3 AAAUCG 706 hsa-miR-518f-3p AAAGCG 5 2 CCUUCU 707 hsa-miR-370-3p CCUGCU 5 6.5 GCAUCA 708 hsa-miR-107/hsa-miR-103a-3p GCAGCA 5 57.4 GGAUUG 709 hsa-miR-122-5p GGAGUG 5 96.6 CCAUCA 710 hsa-miR-338-3p CCAGCA 5 7.3 AAUUUU 711 hsa-miR-409-3p AAUGUU 5 7.8 AAGUCA 712 hsa-miR-124-3p AAGGCA 5 2.7 GAGUUA 713 hsa-let-7d-5p/hsa-let-7g-5p/hsa-let-7i-5p/hsa- GAGGUA 5 23.9 let-7f-5p/hsa-let-7e-5p/hsa-let-7a-5p/hsa-let- 7b-5p/hsa-let-7c-5p AGUUCA 714 hsa-miR-130b-3p/hsa-miR-301a-3p/hsa-miR- AGUGCA 5 4 130a-3p/hsa-miR-301b-3p AGUUCU 715 hsa-miR-512-3p AGUGCU 5 2.7 AACUGA 716 hsa-miR-191-5p AACGGA 5 6.8 ACUUCA 717 hsa-miR-509-3-5p ACUGCA 5 38.2 AUUUCA 718 hsa-miR-92a-3p/hsa-miR-92b-3p/hsa-miR- AUUGCA 5 41 363-3p/hsa-miR-25-3p/hsa-miR-32-5p AAGUUG 719 hsa-miR-18a-5p AAGGUG 5 7 AUGUCA 720 hsa-miR-183-5p AUGGCA 5 8.1 GCUUGU 721 hsa-miR-138-5p GCUGGU 5 2.8 CUCUGC 722 hsa-miR-1307-3p CUCGGC 5 2.1 GAGUGG 723 hsa-miR-423-5p GAGGGG 5 3.5 UAAUAC 724 hsa-miR-499a-5p/hsa-miR-208a-3p UAAGAC 5 4.5 CUGUAC 725 hsa-miR-378a-3p CUGGAC 5 8.8 AAAUAA 726 hsa-miR-186-5p AAAGAA 5 3.9 UUUUCA 727 hsa-miR-450b-5p UUUGCA 5 2.3 UUUUCG 728 hsa-miR-450a-5p UUUGCG 5 5.1 GUAUUG 729 hsa-miR-142-3p GUAGUG 5 2.5 ACAUUA 730 hsa-miR-101-3p/hsa-miR-144-3p ACAGUA 5 8.9 AAAUCU 731 hsa-miR-320a AAAGCU 5 6.6 CCAUUG 732 hsa-miR-199b-5p/hsa-miR-199a-5p CCAGUG 5 9.2 UAGUGC 733 hsa-miR-1296-5p UAGGGC 5 2.9 GGAUAG 734 hsa-miR-185-5p GGAGAG 5 2.6 AUGUCU 735 hsa-miR-135a-5p AUGGCU 5 3.1 AGUAUA 736 hsa-miR-411-5p AGUAGA 6 2.5 UCCAUU 737 hsa-miR-145-5p UCCAGU 6 4.2 UCAAUU 738 hsa-miR-26a-5p UCAAGU 6 6.1 ACUGUC 739 hsa-miR-193b-3p ACUGGC 6 2.1 GGCAUU 740 hsa-miR-34c-5p GGCAGU 6 7.1 GGUAUA 741 hsa-miR-379-5p GGUAGA 6 2.1 CCCUUA 742 hsa-miR-125b-5p CCCUGA 6 3.8 CCUGUC 743 hsa-miR-221-5p CCUGGC 6 3.1 UCUUUA 744 hsa-miR-526b-5p UCUUGA 6 4.4 GGAAUA 745 hsa-miR-7-5p GGAAGA 6 4.4 AGCAUC 746 hsa-miR-16-5p/hsa-miR-15b-5p/hsa-miR-424- AGCAGC 6 4.7 5p/hsa-miR-15a-5p UAAAUC 747 hsa-miR-9-3p UAAAGC 6 4.6 GGGUUG 748 hsa-miR-363-5p GGGUGG 6 3 AUCUUG 749 hsa-miR-1298-3p AUCUGG 6 11 GAUUUG 750 hsa-miR-509-3p GAUUGG 6 4.4 GCGGUG 751 hsa-miR-744-5p GCGGGG 6 2.1 CAGUUC 752 hsa-miR-148a-3p CAGUGC 6 36.8 AAGUUC 753 hsa-miR-302a-3p AAGUGC 6 9.8 UAGGUC 754 hsa-miR-1296-5p UAGGGC 6 3.3 GAGGUG 755 hsa-miR-423-5p GAGGGG 6 3.7 CUUUUG 756 hsa-miR-9-5p CUUUGG 6 40 GCUGUU 757 hsa-miR-138-5p GCUGGU 6 3.7 AGCUUC 758 hsa-miR-22-3p AGCUGC 6 47 ACUAUA 759 hsa-miR-28-3p ACUAGA 6 3.4 GAUUUU 760 hsa-miR-508-3p GAUUGU 6 8.4 UAUUUC 761 hsa-miR-137 UAUUGC 6 7.2 GGGGUA 762 hsa-miR-5010-5p GGGGGA 6 2 UCUAUA 763 hsa-miR-523-5p UCUAGA 6 2.5 AACGUA 764 hsa-miR-191-5p AACGGA 6 5 CACAUU 765 hsa-miR-128-3p CACAGU 6 2.8 CCAGUU 766 hsa-miR-199a-5p/hsa-miR-199b-5p CCAGUG 7 6.4 CCACUU 767 hsa-miR-181a-2-3p CCACUG 7 5.3 UCACAU 768 hsa-miR-27a-3p/hsa-miR-27b-3p UCACAG 7 5.4 UAUACU 769 hsa-let-7d-3p UAUACG 7 3.3 UUUUUU 770 hsa-miR-129-5p UUUUUG 7 2.2 UGUGCU 771 hsa-miR-210-3p UGUGCG 7 3.6 GAAUUU 772 hsa-miR-219a-2-3p GAAUUG 7 9.2 AAAACU 773 hsa-miR-424-3p AAAACG 7 2.7 AAGGUU 774 hsa-miR-18a-5p AAGGUG 7 5.1 UGAAAU 775 hsa-miR-488-3p UGAAAG 7 2.6 GGAAUU 776 hsa-miR-1-3p GGAAUG 7 3.8 CCAUCU 777 hsa-miR-181a-3p CCAUCG 7 2.4 CAGUAU 778 hsa-miR-199b-3p CAGUAG 7 7.9 ACCCUU 779 hsa-miR-10a-5p ACCCUG 7 3 AGGUAU 780 hsa-miR-196b-5p AGGUAG 7 2.1 GGUUGU 781 hsa-miR-92a-1-5p GGUUGG 7 2.2 AGACGU 782 hsa-miR-483-5p AGACGG 7 2.2 CUGCAU 783 hsa-miR-17-3p CUGCAG 7 2.1 GGGUGU 784 hsa-miR-363-5p GGGUGG 7 2 CUUUGU 785 hsa-miR-9-5p CUUUGG 7 63.3 AAACCU 786 hsa-miR-1537-3p AAACCG 7 2 AAAGUU 787 hsa-miR-106b-5p/hsa-miR-20a-5p/hsa-miR- AAAGUG 7 8.4 17-5p/hsa-miR-93-5p GAGAUU 788 hsa-miR-143-3p GAGAUG 7 33.4 UUUCAU 789 hsa-miR-30a-3p/hsa-miR-30e-3p UUUCAG 7 8.7 GCUCGU 790 hsa-miR-423-3p GCUCGG 7 2.1 AUCUGU 791 hsa-miR-1298-3p AUCUGG 7 3.7 CGACCU 792 hsa-miR-1307-5p CGACCG 7 3.5 GAGGGU 793 hsa-miR-423-5p GAGGGG 7 2.8 GGAGUU 794 hsa-miR-122-5p GGAGUG 7 53.7 UUAUCAU 795 hsa-miR-374a-3p UUAUCAG 8 4.9 GGUGCGU 796 hsa-miR-675-5p GGUGCGG 8 2 GGUUGGU 797 hsa-miR-92a-1-5p GGUUGGG 8 2.1 CGACCGU 798 hsa-miR-1307-5p CGACCGG 8 6.7 AGCAGCU 799 hsa-miR-503-5p AGCAGCG 8 15.3 GGCUCAU 800 hsa-miR-24-3p GGCUCAG 8 4.9 UAUAAAU 801 hsa-miR-340-5p UAUAAAG 8 3.6 ACUGCAU 802 hsa-miR-509-3-5p ACUGCAG 8 6.3 GGCAGUU 803 hsa-miR-34a-5p/hsa-miR-34c-5p GGCAGUG 8 4.2 UCUUGAU 804 hsa-miR-526b-5p UCUUGAG 8 6.3 UGAAAUU 805 hsa-miR-203a-3p UGAAAUG 8 18.8 UGCAUAU 806 hsa-miR-153-3p UGCAUAG 8 3.4 UAAGACU 807 hsa-miR-208a-3p UAAGACG 8 9.8 AAUACUU 808 hsa-miR-200b-3p/hsa-miR-200c-3p AAUACUG 8 15.6 UCUAGAU 809 hsa-miR-518f-5p/hsa-miR-523-5p UCUAGAG 8 6.4 ACUAUAU 810 hsa-miR-625-3p ACUAUAG 8 2.9 GUAACAU 811 hsa-miR-194-5p GUAACAG 8 12.9 UGUACAU 812 hsa-let-7g-3p UGUACAG 8 2.2 AUCUGGU 813 hsa-miR-1298-3p AUCUGGG 8 2.1 ACUCUGU 814 hsa-miR-514a-5p ACUCUGG 8 2.6 AGACGGU 815 hsa-miR-483-5p AGACGGG 8 12 CGUACCU 816 hsa-miR-126-3p CGUACCG 8 13.7 CACAGUU 817 hsa-miR-128-3p CACAGUG 8 12.1 AUACAAU 818 hsa-miR-381-3p AUACAAG 8 7.1 AAAGCUU 819 hsa-miR-320a AAAGCUG 8 4.1 UCUCAAU 820 hsa-miR-513c-5p/hsa-miR-514b-5p UCUCAAG 8 4.3 GCUGGUU 821 hsa-miR-138-5p GCUGGUG 8 3.4 UCCAGAU 822 hsa-miR-520a-5p UCCAGAG 8 4.4 CCCUGAU 823 hsa-miR-125b-5p/hsa-miR-125a-5p CCCUGAG 8 21.3 AACACUU 824 hsa-miR-141-3p AACACUG 8 2.4 UGCCCUU 825 hsa-miR-874-3p UGCCCUG 8 3.5 UCCUAUU 826 hsa-miR-202-5p UCCUAUG 8 22.5 ACCACAU 827 hsa-miR-140-3p ACCACAG 8 3.2 CCCCCAU 828 hsa-miR-361-3p CCCCCAG 8 2 UCACAAU 829 hsa-miR-513b-5p UCACAAG 8 2.5 GUGACUU 830 hsa-miR-134-5p GUGACUG 8 2.9 UGCAUUU 831 hsa-miR-33a-5p UGCAUUG 8 2.1 AGUGCUU 832 hsa-miR-512-3p AGUGCUG 8 2.7 GAGGUAU 833 hsa-let-7a-5p/hsa-let-7c-5p/hsa-let-7b-5p/hsa- GAGGUAG 8 36.5 let-7d-5p/hsa-let-7f-5p/hsa-let-7e-5p/hsa-let- 7i-5p/hsa-let-7g-5p UUGUUCU 834 hsa-miR-375 UUGUUCG 8 25.5 AUCAUCU 835 hsa-miR-136-3p AUCAUCG 8 6.3 ACUCCAU 836 hsa-miR-508-5p ACUCCAG 8 10.6 AAGGCACU 837 hsa-miR-124-3p AAGGCACG 9 2.7 UCCCUUUU 838 hsa-miR-204-5p/hsa-miR-211-5p UCCCUUUG 9 2.6 UGUGCGUU 839 hsa-miR-210-3p UGUGCGUG 9 3.8 GAGAACUU 840 hsa-miR-146a-5p/hsa-miR-146b-5p GAGAACUG 9 5.4 GAUUGUAU 841 hsa-miR-508-3p GAUUGUAG 9 4.9 UCACAUUU 842 hsa-miR-23a-3p UCACAUUG 9 6.1 GAAUUGUU 843 hsa-miR-219a-2-3p GAAUUGUG 9 11.4 AACACCAU 844 hsa-miR-21-3p AACACCAG 9 2.7 GGAGUGUU 845 hsa-miR-122-5p GGAGUGUG 9 42.5 UAGAGGAU 846 hsa-miR-877-5p UAGAGGAG 9 2.3 UUUUUGCU 847 hsa-miR-129-5p UUUUUGCG 9 5.8 UCUUGAGU 848 hsa-miR-526b-5p UCUUGAGG 9 4.1 CUUAAACU 849 hsa-miR-302a-5p CUUAAACG 9 13 AUGCCUUU 850 hsa-miR-532-5p AUGCCUUG 9 3.8 UCCAGAGU 851 hsa-miR-520a-5p UCCAGAGG 9 2 CUACAGUU 852 hsa-miR-139-5p CUACAGUG 9 2 ACCCGUAU 853 hsa-miR-99a-5p/hsa-miR-100-5p/hsa-miR-99b-5p ACCCGUAG 9 34.8 AAAGCUGU 854 hsa-miR-320a AAAGCUGG 9 2.6 AAUCUCAU 855 hsa-miR-216a-5p AAUCUCAG 9 5.5 CGGAUCCU 856 hsa-miR-127-3p CGGAUCCG 9 4.3 CGGGUUAU 857 hsa-miR-125b-1-3p CGGGUUAG 9 4.7 AUACAAGU 858 hsa-miR-381-3p AUACAAGG 9 2 UCACAGUU 859 hsa-miR-27a-3p/hsa-miR-27b-3p UCACAGUG 9 5.6 GGGGGAUU 860 hsa-miR-5010-5p GGGGGAUG 9 2.5 UGCCCUAU 861 hsa-miR-3157-3p UGCCCUAG 9 2 CUGCAGUU 862 hsa-miR-17-3p CUGCAGUG 9 2.4 UCUAGAGU 863 hsa-miR-523-5p UCUAGAGG 9 2.8
[0372] Through the miRNA sequencing variant analyses in the example, variant miRNAs recognizing a non-canonical G:A wobble seed target as a canonical target in various cancer patients were identified, and therefore, when the present invention (miRNA-GU) for modifying a miRNA sequence to recognize a non-canonical G:A wobble seed site as a canonical seed site is applied, a total of 335 sequences (sequence (G>U)) were made (Table 4), thereby completing the technology of exhibiting only the function of suppressing a non-canonical target of miRNA.
INDUSTRIAL APPLICABILITY
[0373] When an interference-inducing nucleic acid according to the present invention is used, a biological function exhibited by suppressing a non-canonical target gene of conventional miRNA may be effectively improved, or a part of the functions of the convention miRNA, that is, only a biological function exhibited by suppressing a non-canonical target gene may be selectively exhibited. In addition, cell cycling, differentiation, dedifferentiation, morphology, migration, proliferation or apoptosis may be regulated by the interference-inducing nucleic acid, and thus it is expected to be used in various fields such as pharmaceuticals, cosmetics, etc.