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  4. Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System

Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System

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Author(s)
Makarova, Kira S.
•
van der Oost, John
•
Koonin, Eugene V.
•
Zetsche, Bernd
•
Gootenberg, Jonathan S
•
Abudayyeh, Omar Osama
•
Slaymaker, Ian
•
Essletzbichler, Patrick
•
Volz, Sara E.
•
Joung, Julia
more
Date Issued
September 2015
Journal
Cell
Publisher
Elsevier
Citation
Zetsche, Bernd et al. “Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System.” Cell 163.3 (2015): 759–771.
Version
Author's final manuscript
Abstract
The microbial adaptive immune system CRISPR mediates defense against foreign genetic elements through two classes of RNA-guided nuclease effectors. Class 1 effectors utilize multiprotein complexes, whereas Class 2 effectors rely on single-component effector proteins such as the well-characterized Cas9. Here we report characterization of Cpf1, a putative Class 2 CRISPR effector. We demonstrate that Cpf1 mediates robust DNA interference with features distinct from Cas9. Cpf1 is a single RNA-guided endonuclease lacking tracrRNA, and it utilizes a T-rich protospacer adjacent motif. Moreover, Cpf1 cleaves DNA via a staggered DNA double stranded break. Out of 16 Cpf1-family proteins, we identified two candidate enzymes, from Acidominococcus and Lachnospiraceae, with efficient genome editing activity in human cells. Identifying this mechanism of interference broadens our understanding of CRISPR-Cas systems and advances their genome editing applications.
MIT Department
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
McGovern Institute for Brain Research at MIT
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
http://creativecommons.org/licenses/by-nc-nd/4.0/
Persistent DSpace Link
http://hdl.handle.net/1721.1/105747
DOI of Published Version
https://doi.org/10.1016/j.cell.2015.09.038
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