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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.

Characterization of CoCas9 nuclease from Capnocytophaga ochracea