Characterization of CoCas9 nuclease from Capnocytophaga ochracea

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 Cap...

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Main Authors: A. Vasileva, P. Selkova, A. Arseniev, M. Abramova, N. Shcheglova, O. Musharova, I. Mizgirev, T. Artamonova, M. Khodorkovskii, K. Severinov, I. Fedorova
Format: Article
Language:English
Published: Taylor & Francis Group 2023-12-01
Series:RNA Biology
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Online Access:https://www.tandfonline.com/doi/10.1080/15476286.2023.2256578
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author A. Vasileva
P. Selkova
A. Arseniev
M. Abramova
N. Shcheglova
O. Musharova
I. Mizgirev
T. Artamonova
M. Khodorkovskii
K. Severinov
I. Fedorova
author_facet A. Vasileva
P. Selkova
A. Arseniev
M. Abramova
N. Shcheglova
O. Musharova
I. Mizgirev
T. Artamonova
M. Khodorkovskii
K. Severinov
I. Fedorova
author_sort A. Vasileva
collection DOAJ
description 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.
format Article
id doaj-art-08bbbcf3e60448a3b5a361c6aeec2e26
institution DOAJ
issn 1547-6286
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language English
publishDate 2023-12-01
publisher Taylor & Francis Group
record_format Article
series RNA Biology
spelling doaj-art-08bbbcf3e60448a3b5a361c6aeec2e262025-08-20T03:12:56ZengTaylor & Francis GroupRNA Biology1547-62861555-85842023-12-0120175075910.1080/15476286.2023.2256578Characterization of CoCas9 nuclease from Capnocytophaga ochraceaA. Vasileva0P. Selkova1A. Arseniev2M. Abramova3N. Shcheglova4O. Musharova5I. Mizgirev6T. Artamonova7M. Khodorkovskii8K. Severinov9I. Fedorova10Center of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaComplex of NBICS Technologies, National Research Center “Kurchatov Institute”, Moscow, RussiaLaboratory of Carcinogenesis and Aging, N.N. Petrov National Medical Research Center of Oncology, St. Petersburg, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaComplex of NBICS Technologies, National Research Center “Kurchatov Institute”, Moscow, RussiaCenter of Nanobiotechnology, Peter the Great St. Petersburg Polytechnic University, St. Petersburg, RussiaCas9 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.https://www.tandfonline.com/doi/10.1080/15476286.2023.2256578CoCas9Cas9Capnocytophaga ochraceaCRISPR-CasRNA-guided nuclease
spellingShingle A. Vasileva
P. Selkova
A. Arseniev
M. Abramova
N. Shcheglova
O. Musharova
I. Mizgirev
T. Artamonova
M. Khodorkovskii
K. Severinov
I. Fedorova
Characterization of CoCas9 nuclease from Capnocytophaga ochracea
RNA Biology
CoCas9
Cas9
Capnocytophaga ochracea
CRISPR-Cas
RNA-guided nuclease
title Characterization of CoCas9 nuclease from Capnocytophaga ochracea
title_full Characterization of CoCas9 nuclease from Capnocytophaga ochracea
title_fullStr Characterization of CoCas9 nuclease from Capnocytophaga ochracea
title_full_unstemmed Characterization of CoCas9 nuclease from Capnocytophaga ochracea
title_short Characterization of CoCas9 nuclease from Capnocytophaga ochracea
title_sort characterization of cocas9 nuclease from capnocytophaga ochracea
topic CoCas9
Cas9
Capnocytophaga ochracea
CRISPR-Cas
RNA-guided nuclease
url https://www.tandfonline.com/doi/10.1080/15476286.2023.2256578
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