Cpf1 is a single RNA-guided endonuclease of a Class 2 CRISPR-Cas system

Zetsche, Bernd; Gootenberg, Jonathan S.; Abudayyeh, Omar O.; Slaymaker, Ian M.; Makarova, Kira S.; Essletzbichler, Patrick; Volz, Sara; Joung, Julia; van der Oost, John; Regev, Aviv; Koonin, Eugene V.; Zhang, Feng

doi:10.1016/j.cell.2015.09.038

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Cpf1 is a single RNA-guided endonuclease of a Class 2 CRISPR-Cas system

10 22 2015
By Zetsche, Bernd ; Gootenberg, Jonathan S. ; Abudayyeh, Omar O. ; ...
http://dx.doi.org/10.1016/j.cell.2015.09.038
Source: Cell. 163(3):759-771

[PDF-1.79 MB]

English

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Alternative Title:

Cell
Personal Author:

Zetsche, Bernd ; Gootenberg, Jonathan S. ; Abudayyeh, Omar O. ; Slaymaker, Ian M. ; Makarova, Kira S. ; ... More +

Zetsche, Bernd ; Gootenberg, Jonathan S. ; Abudayyeh, Omar O. ; Slaymaker, Ian M. ; Makarova, Kira S. ; Essletzbichler, Patrick ; Volz, Sara ; Joung, Julia ; van der Oost, John ; Regev, Aviv ; Koonin, Eugene V. ; Zhang, Feng Less -
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.
Subjects:

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Amino Acid Sequence Article CRISPR-Cas Systems Endonucleases Francisella Genetic Engineering HEK293 Cells Humans Molecular Sequence Data Nucleic Acid Conformation RNA, Guide, Kinetoplastida Sequence Alignment
Source:

Cell. 163(3):759-771
Pubmed ID:

26422227
Pubmed Central ID:

PMC4638220
Document Type:

Journal Article
Funding:

DP1 MH100706/MH/NIMH NIH HHSUnited States/ ; R01 MH110049/MH/NIMH NIH HHSUnited States/ ; 5DP1-MH100706/DP/NCCDPHP CDC HHSUnited States/

DP1 MH100706/MH/NIMH NIH HHSUnited States/ ; R01 MH110049/MH/NIMH NIH HHSUnited States/ ; 5DP1-MH100706/DP/NCCDPHP CDC HHSUnited States/ Less -
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CDC Public Access
Main Document Checksum:

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urn:sha256:cc11c5ba40f6c17ab27cfcdf9f8c751323492b06f1b5620c927c0791e2bf3264
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https://stacks.cdc.gov/view/cdc/37605/cdc_37605_DS1.pdf
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