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Nat Commun ; 13(1): 7602, 2022 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-36522342

RESUMO

Programmable, RNA-guided nucleases are diverse enzymes that have been repurposed for biotechnological applications. However, to further expand the therapeutic application of these tools there is a need for targetable systems that are small enough to be delivered efficiently. Here, we mined an extensive genome-resolved metagenomics database and identified families of uncharacterized RNA-guided, compact nucleases (between 450 and 1,050 aa). We report that Cas9d, a new CRISPR type II subtype, contains Zinc-finger motifs and high arginine content, features that we also found in nucleases related to HEARO effectors. These enzymes exhibit diverse biochemical characteristics and are broadly targetable. We show that natural Cas9d enzymes are capable of genome editing in mammalian cells with >90% efficiency, and further engineered nickase variants into the smallest base editors active in E. coli and human cells. Their small size, broad targeting potential, and translatability suggest that Cas9d and HEARO systems will enable a variety of genome editing applications.


Assuntos
Escherichia coli , Edição de Genes , Animais , Humanos , Escherichia coli/genética , Escherichia coli/metabolismo , Endonucleases/genética , Endonucleases/metabolismo , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Ribonucleases/genética , RNA , Sistemas CRISPR-Cas/genética , Mamíferos/genética
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