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DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system
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 restrict...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7038990/ https://www.ncbi.nlm.nih.gov/pubmed/31943070 http://dx.doi.org/10.1093/nar/gkz1225 |
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author | Fedorova, Iana Arseniev, Anatolii Selkova, Polina Pobegalov, Georgii Goryanin, Ignatiy Vasileva, Aleksandra Musharova, Olga Abramova, Marina Kazalov, Maksim Zyubko, Tatyana Artamonova, Tatyana Artamonova, Daria Shmakov, Sergey Khodorkovskii, Mikhail Severinov, Konstantin |
author_facet | Fedorova, Iana Arseniev, Anatolii Selkova, Polina Pobegalov, Georgii Goryanin, Ignatiy Vasileva, Aleksandra Musharova, Olga Abramova, Marina Kazalov, Maksim Zyubko, Tatyana Artamonova, Tatyana Artamonova, Daria Shmakov, Sergey Khodorkovskii, Mikhail Severinov, Konstantin |
author_sort | Fedorova, Iana |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7038990 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-70389902020-03-02 DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system Fedorova, Iana Arseniev, Anatolii Selkova, Polina Pobegalov, Georgii Goryanin, Ignatiy Vasileva, Aleksandra Musharova, Olga Abramova, Marina Kazalov, Maksim Zyubko, Tatyana Artamonova, Tatyana Artamonova, Daria Shmakov, Sergey Khodorkovskii, Mikhail Severinov, Konstantin Nucleic Acids Res Nucleic Acid Enzymes 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. Oxford University Press 2020-02-28 2020-01-16 /pmc/articles/PMC7038990/ /pubmed/31943070 http://dx.doi.org/10.1093/nar/gkz1225 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Fedorova, Iana Arseniev, Anatolii Selkova, Polina Pobegalov, Georgii Goryanin, Ignatiy Vasileva, Aleksandra Musharova, Olga Abramova, Marina Kazalov, Maksim Zyubko, Tatyana Artamonova, Tatyana Artamonova, Daria Shmakov, Sergey Khodorkovskii, Mikhail Severinov, Konstantin DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system |
title | DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system |
title_full | DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system |
title_fullStr | DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system |
title_full_unstemmed | DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system |
title_short | DNA targeting by Clostridium cellulolyticum CRISPR–Cas9 Type II-C system |
title_sort | dna targeting by clostridium cellulolyticum crispr–cas9 type ii-c system |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7038990/ https://www.ncbi.nlm.nih.gov/pubmed/31943070 http://dx.doi.org/10.1093/nar/gkz1225 |
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