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DNA targeting by Clostridium cellulolyticum CRISPR-Cas9 Type II-C system.

ABSTRACT: Type II CRISPR-Cas9 RNA-guided nucleases are widely used for genome engineering. Type II-A SpCas9 protein from Streptococcus pyogenes is the most investigated and highly used enzyme of its class. Nevertheless, it has some drawbacks, including a relatively big size, imperfect specificity and restriction to DNA targets flanked by an NGG PAM sequence. Cas9 orthologs from other bacterial species may provide a rich and largely untapped source of biochemical diversity, which can help to overcome the limitations of SpCas9. Here, we characterize CcCas9, a Type II-C CRISPR nuclease from Clostridium cellulolyticum H10. We show that CcCas9 is an active endonuclease of comparatively small size that recognizes a novel two-nucleotide PAM sequence. The CcCas9 can potentially broaden the existing scope of biotechnological applications of Cas9 nucleases and may be particularly advantageous for genome editing of C. cellulolyticum H10, a bacterium considered to be a promising biofuel producer.

SUBMITTER: Fedorova I

PROVIDER: S-EPMC7038990 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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DNA targeting by Clostridium cellulolyticum CRISPR-Cas9 Type II-C system.

Fedorova Iana I Arseniev Anatolii A Selkova Polina P Pobegalov Georgii G Goryanin Ignatiy I Vasileva Aleksandra A Musharova Olga O Abramova Marina M Kazalov Maksim M Zyubko Tatyana T Artamonova Tatyana T Artamonova Daria D Shmakov Sergey S Khodorkovskii Mikhail M Severinov Konstantin K

Nucleic acids research 20200201 4

Type II CRISPR-Cas9 RNA-guided nucleases are widely used for genome engineering. Type II-A SpCas9 protein from Streptococcus pyogenes is the most investigated and highly used enzyme of its class. Nevertheless, it has some drawbacks, including a relatively big size, imperfect specificity and restriction to DNA targets flanked by an NGG PAM sequence. Cas9 orthologs from other bacterial species may provide a rich and largely untapped source of biochemical diversity, which can help to overcome the l ...[more]

PMID: 31943070

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Dataset Information

DNA targeting by Clostridium cellulolyticum CRISPR-Cas9 Type II-C system.

Publications

DNA targeting by Clostridium cellulolyticum CRISPR-Cas9 Type II-C system.

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