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

Cell, 2015-10, Vol.163 (3), p.759-771 [Peer Reviewed Journal]

2015 Elsevier Inc. ;Copyright © 2015 Elsevier Inc. All rights reserved. ;Wageningen University & Research ;ISSN: 0092-8674 ;EISSN: 1097-4172 ;DOI: 10.1016/j.cell.2015.09.038 ;PMID: 26422227

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Title:
Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
Author: Zetsche, Bernd ; Gootenberg, Jonathan S. ; Abudayyeh, Omar O. ; Slaymaker, Ian M. ; Makarova, Kira S. ; Essletzbichler, Patrick ; Volz, Sara E. ; Joung, Julia ; van der Oost, John ; Regev, Aviv ; Koonin, Eugene V. ; Zhang, Feng
Subjects: Amino Acid Sequence ; CRISPR-Cas Systems ; Endonucleases - chemistry ; Endonucleases - genetics ; Francisella - enzymology ; Francisella - genetics ; Genetic Engineering - methods ; HEK293 Cells ; Humans ; Laboratorium voor Microbiologie ; Microbiological Laboratory ; Microbiologie ; Microbiology ; Molecular Sequence Data ; Nucleic Acid Conformation ; RNA, Guide, CRISPR-Cas Systems - genetics ; Sequence Alignment ; VLAG
Is Part Of: Cell, 2015-10, Vol.163 (3), p.759-771
Description: The microbial adaptive immune system CRISPR mediates defense against foreign genetic elements through two classes of RNA-guided nuclease effectors. Class 1 effectors utilize multi-protein 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 Acidaminococcus 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. [Display omitted] •CRISPR-Cpf1 is a class 2 CRISPR system•Cpf1 is a CRISPR-associated two-component RNA-programmable DNA nuclease•Targeted DNA is cleaved as a 5-nt staggered cut distal to a 5′ T-rich PAM•Two Cpf1 orthologs exhibit robust nuclease activity in human cells Cpf1 is a RNA-guided DNA nuclease that provides immunity in bacteria and can be adapted for genome editing in mammalian cells.
Publisher: United States: Elsevier Inc
Language: English
Identifier: ISSN: 0092-8674
EISSN: 1097-4172
DOI: 10.1016/j.cell.2015.09.038
PMID: 26422227
Source: Cell Press Free Archives
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Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System

2015 Elsevier Inc. ;Copyright © 2015 Elsevier Inc. All rights reserved. ;Wageningen University & Research ;ISSN: 0092-8674 ;EISSN: 1097-4172 ;DOI: 10.1016/j.cell.2015.09.038 ;PMID: 26422227

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