DNA targeting by Clostridium cellulolyticum CRISPR-Cas9 Type II-C system.
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
28 02 2020
28 02 2020
Historique:
accepted:
20
12
2019
revised:
16
12
2019
received:
20
07
2019
pubmed:
17
1
2020
medline:
19
3
2020
entrez:
17
1
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31943070
pii: 5707196
doi: 10.1093/nar/gkz1225
pmc: PMC7038990
doi:
Substances chimiques
RNA, Guide
0
DNA
9007-49-2
CRISPR-Associated Protein 9
EC 3.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2026-2034Informations de copyright
© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
Références
Nat Biotechnol. 2013 Mar;31(3):233-9
pubmed: 23360965
FEMS Microbiol Rev. 2005 Sep;29(4):741-64
pubmed: 16102601
Mol Microbiol. 2010 Sep;77(6):1367-79
pubmed: 20624226
Nat Commun. 2017 Nov 10;8(1):1424
pubmed: 29127284
Nucleic Acids Res. 2014 Feb;42(4):2577-90
pubmed: 24270795
Science. 2012 Aug 17;337(6096):816-21
pubmed: 22745249
Nat Commun. 2017 Feb 21;8:14500
pubmed: 28220790
Science. 2013 Feb 15;339(6121):819-23
pubmed: 23287718
Methods. 2018 Jul 1;143:48-57
pubmed: 29486239
Sci Rep. 2016 Jan 22;6:19452
pubmed: 26797514
Bioinformatics. 2009 Jul 15;25(14):1754-60
pubmed: 19451168
Front Microbiol. 2017 Sep 07;8:1744
pubmed: 28936208
Nature. 2015 Apr 9;520(7546):186-91
pubmed: 25830891
Mol Cell. 2016 Apr 7;62(1):137-47
pubmed: 27041224
Sci Rep. 2016 May 09;6:25666
pubmed: 27157668
Appl Environ Microbiol. 2015 Jul;81(13):4423-31
pubmed: 25911483
Mol Ther. 2016 Mar;24(3):645-54
pubmed: 26782639
Mol Cell. 2013 May 23;50(4):488-503
pubmed: 23706818
Nat Commun. 2017 May 04;8:15179
pubmed: 28469274