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Characterization of CoCas9 nuclease from Capnocytophaga ochracea.

ABSTRACT: Cas9 nucleases are widely used for genome editing and engineering. Cas9 enzymes encoded by CRISPR-Cas defence systems of various prokaryotic organisms possess different properties such as target site preferences, size, and DNA cleavage efficiency. Here, we biochemically characterized CoCas9 from Capnocytophaga ochracea, a bacterium that inhabits the oral cavity of humans and contributes to plaque formation on teeth. CoCas9 recognizes a novel 5'-NRRWC-3' PAM and efficiently cleaves DNA in vitro. Functional characterization of CoCas9 opens ways for genetic engineering of C. ochracea using its endogenous CRISPR-Cas system. The novel PAM requirement makes CoCas9 potentially useful in genome editing applications.

SUBMITTER: Vasileva A

PROVIDER: S-EPMC10521337 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Characterization of CoCas9 nuclease from Capnocytophaga ochracea.

Vasileva A A Selkova P P Arseniev A A Abramova M M Shcheglova N N Musharova O O Mizgirev I I Artamonova T T Khodorkovskii M M Severinov K K Fedorova I I

RNA biology 20230101 1

Cas9 nucleases are widely used for genome editing and engineering. Cas9 enzymes encoded by CRISPR-Cas defence systems of various prokaryotic organisms possess different properties such as target site preferences, size, and DNA cleavage efficiency. Here, we biochemically characterized CoCas9 from Capnocytophaga ochracea, a bacterium that inhabits the oral cavity of humans and contributes to plaque formation on teeth. CoCas9 recognizes a novel 5'-NRRWC-3' PAM and efficiently cleaves DNA ...[more]

PMID: 37743659

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