Described are compositions and methods for altering mutations associated with Rett Syndrome (RETT). Provided herein are compositions and methods of using base editors (e.g., ABE8) comprising a polynucleotide programmable nucleotide binding domain and a nucleobase editing domain in conjunction with a guide polynucleotide. Also provided herein are base editor systems for editing nucleobases of target nucleotide sequences.

Described are compositions and methods for altering mutations associated with Rett Syndrome (RETT). Provided herein are compositions and methods of using base editors (e.g., ABE8) comprising a polynucleotide programmable nucleotide binding domain and a nucleobase editing domain in conjunction with a guide polynucleotide. Also provided herein are base editor systems for editing nucleobases of target nucleotide sequences.

The invention relates to fusion target-binding proteins, such as chimeric antigen receptors (CARs), that comprise a target binding moiety, an intracellular signalling region, and an arginase domain. These proteins confer advantages that include improved cell killing and increased proliferation. The invention also relates to nucleic acids encoding the fusion target-binding proteins and cells expressing such proteins. The invention relates to pharmaceutical compositions, medical uses, and methods of treatment, all using the fusion target-binding proteins, cells, or nucleic acids disclosed. The medical uses and methods of treatment are of particular benefit in cancer therapy.

The invention relates to fusion target-binding proteins, such as chimeric antigen receptors (CARs), that comprise a target binding moiety, an intracellular signalling region, and an arginase domain. These proteins confer advantages that include improved cell killing and increased proliferation. The invention also relates to nucleic acids encoding the fusion target-binding proteins and cells expressing such proteins. The invention relates to pharmaceutical compositions, medical uses, and methods of treatment, all using the fusion target-binding proteins, cells, or nucleic acids disclosed. The medical uses and methods of treatment are of particular benefit in cancer therapy.

The present invention discloses base editing systems for mutating a base C to A and a base C to G and applications thereof. The base editing system for mutating C to A disclosed in the present invention includes cytosine deaminase AID and nCas9 nuclease or includes cytosine deaminase AID, nCas9 nuclease and uracil DNA glycosidase; the base editing system for mutating C to G of the present invention includes cytosine deaminase APOBEC, nCas9 nuclease and uracil DNA glycosidase. The experiments show that a combination of the three base editing systems for mutating C to A, C to T and A to G can realize a mutation of A, T, C or G to any base in both prokaryotes and eukaryotes.

The present invention discloses base editing systems for mutating a base C to A and a base C to G and applications thereof. The base editing system for mutating C to A disclosed in the present invention includes cytosine deaminase AID and nCas9 nuclease or includes cytosine deaminase AID, nCas9 nuclease and uracil DNA glycosidase; the base editing system for mutating C to G of the present invention includes cytosine deaminase APOBEC, nCas9 nuclease and uracil DNA glycosidase. The experiments show that a combination of the three base editing systems for mutating C to A, C to T and A to G can realize a mutation of A, T, C or G to any base in both prokaryotes and eukaryotes.

The present invention provides chimeric ectolysins useful for selective suppression of certain targeted bacterial species while having little to no effect on closely related non-targeted bacterial species. Specifically, the disclosure provides polypeptides for selective suppression of growth of Staphylococcus aureus and/or S. hominis but not S. epidermidis. Compositions and methods for selective suppression of target bacterial species are also provided.

The type V-U1 system from the bacterium Mycobacterium mucogenicum CCH10-A2 (Mmu) has a nuclease which binds dsDNA but it does not cleave it. Additionally, after dsDNA binding by the nuclease an RuvC-dependent interference of nascent transcript (mRNA) takes place and this mechanism has not been described before for any CRISPR system. CRISPR based gene manipulation can therefore use CRISPR endonucleases from the Type V-U1 system from Mycobacterium mucogenicum, including variant and modified endonucleases, so as to provide for methods of expression control and gene editing in cells of any living organism or of any nucleic acid in vitro.

The type V-U1 system from the bacterium Mycobacterium mucogenicum CCH10-A2 (Mmu) has a nuclease which binds dsDNA but it does not cleave it. Additionally, after dsDNA binding by the nuclease an RuvC-dependent interference of nascent transcript (mRNA) takes place and this mechanism has not been described before for any CRISPR system. CRISPR based gene manipulation can therefore use CRISPR endonucleases from the Type V-U1 system from Mycobacterium mucogenicum, including variant and modified endonucleases, so as to provide for methods of expression control and gene editing in cells of any living organism or of any nucleic acid in vitro.

The present invention provides a nitrilase mutant and application thereof in the synthesis of 1-cyanocyclohexyl acetic acid, the nitrilase mutant is obtained by mutating one or two of the amino acids at position 180 and 205 of the amino acid sequence shown in SEQ ID No. 2. In the present invention, by semi-rational design and protein molecular modification, the specific enzyme activity of the nitrilase double mutant AcN-G180D/A205C was increased by up to 1.6 folds, and the conversion rate>99%. And the reaction time was shortened to a quarter of the original using the recombinant Escherichia coli containing the nitrilase mutant to hydrolyze 1-cyanocyclohexylacetonitrile at high temperature (50° C.). Therefore, the mutants obtained by the present invention have a good application prospect in efficiently catalyzing 1-cyanocyclohexylacetonitrile to synthesize gabapentin intermediate, 1-cyanocyclohexyl acetic acid.