Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems
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nihms-729607.pdf
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1.29 MB
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Checksum (MD5)
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Author(s)
Shmakov, Sergey
Makarova, Kira S.
Wolf, Yuri I.
Semenova, Ekaterina
Minakhin, Leonid
Severinov, Konstantin
Koonin, Eugene V.
Abudayyeh, Omar Osama
Gootenberg, Jonathan S
Joung, Julia
Date Issued
October 2015
Journal
Molecular Cell
Publisher
Elsevier
Citation
Shmakov, Sergey et al. “Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems.” Molecular Cell 60, 3 (November 2015): 385–397 © 2015 Elsevier
Version
Author's final manuscript
Abstract
Microbial CRISPR-Cas systems are divided into Class 1, with multisubunit effector complexes, and Class 2, with single protein effectors. Currently, only two Class 2 effectors, Cas9 and Cpf1, are known. We describe here three distinct Class 2 CRISPR-Cas systems. The effectors of two of the identified systems, C2c1 and C2c3, contain RuvC-like endonuclease domains distantly related to Cpf1. The third system, C2c2, contains an effector with two predicted HEPN RNase domains. Whereas production of mature CRISPR RNA (crRNA) by C2c1 depends on tracrRNA, C2c2 crRNA maturation is tracrRNA independent. We found that C2c1 systems can mediate DNA interference in a 5'-PAM-dependent fashion analogous to Cpf1. However, unlike Cpf1, which is a single-RNA-guided nuclease, C2c1 depends on both crRNA and tracrRNA for DNA cleavage. Finally, comparative analysis indicates that Class 2 CRISPR-Cas systems evolved on multiple occasions through recombination of Class 1 adaptation modules with effector proteins acquired from distinct mobile elements.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
McGovern Institute for Brain Research at MIT
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Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/J.MOLCEL.2015.10.008
