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Characterization of CoCas9 nuclease from Capnocytophaga ochracea

Vasileva, A ; Selkova, P LU ; Arseniev, A ; Abramova, M ; Shcheglova, N ; Musharova, O ; Mizgirev, I ; Artamonova, T ; Khodorkovskii, M and Severinov, K , et al. (2023) In RNA Biology 20(1). p.750-759
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.

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publishing date
type
Contribution to journal
publication status
published
keywords
Humans, CRISPR-Associated Protein 9/genetics, CRISPR-Cas Systems, Gene Editing, Capnocytophaga/genetics
in
RNA Biology
volume
20
issue
1
pages
750 - 759
publisher
Taylor & Francis
external identifiers
  • pmid:37743659
ISSN
1547-6286
DOI
10.1080/15476286.2023.2256578
language
English
LU publication?
no
id
0bde0802-ddb7-464c-a1f4-e1a25ada6a84
date added to LUP
2026-01-29 09:51:20
date last changed
2026-01-29 10:04:03
@article{0bde0802-ddb7-464c-a1f4-e1a25ada6a84,
  abstract     = {{<p>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.</p>}},
  author       = {{Vasileva, A and Selkova, P and Arseniev, A and Abramova, M and Shcheglova, N and Musharova, O and Mizgirev, I and Artamonova, T and Khodorkovskii, M and Severinov, K and Fedorova, I}},
  issn         = {{1547-6286}},
  keywords     = {{Humans; CRISPR-Associated Protein 9/genetics; CRISPR-Cas Systems; Gene Editing; Capnocytophaga/genetics}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{750--759}},
  publisher    = {{Taylor & Francis}},
  series       = {{RNA Biology}},
  title        = {{Characterization of CoCas9 nuclease from Capnocytophaga ochracea}},
  url          = {{http://dx.doi.org/10.1080/15476286.2023.2256578}},
  doi          = {{10.1080/15476286.2023.2256578}},
  volume       = {{20}},
  year         = {{2023}},
}