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

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          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 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 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.

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          Author and article information

          Journal
          0413066
          2830
          Cell
          Cell
          Cell
          0092-8674
          1097-4172
          29 September 2015
          25 September 2015
          22 October 2015
          22 October 2016
          : 163
          : 3
          : 759-771
          Affiliations
          [1 ]Broad Institute of MIT and Harvard, Cambridge, MA 02142
          [2 ]McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139
          [3 ]Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139
          [4 ]Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139
          [5 ]Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Sigmund Freud Street 25, 53127 Bonn, Germany
          [6 ]Department of Systems Biology, Harvard Medical School, Boston, MA 02115
          [7 ]National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20894
          [8 ]Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands
          [9 ]Department of Biology, Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139
          Author notes
          []To whom correspondence should be addressed: zhang@ 123456broadinstitute.org (F.Z.)
          [*]

          These authors contributed equally to this work.

          Article
          PMC4638220 PMC4638220 4638220 nihpa725840
          10.1016/j.cell.2015.09.038
          4638220
          26422227
          451e0b21-f7ea-41b3-8d0a-7c7aa06f043b
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