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

Details

• Alternative Title:
  Cell
• Personal Author:
  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
• 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:
  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

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• 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/
• Volume:
  163
• Issue:
  3
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:cc11c5ba40f6c17ab27cfcdf9f8c751323492b06f1b5620c927c0791e2bf3264
• Download URL:
  https://stacks.cdc.gov/view/cdc/37605/cdc_37605_DS1.pdf
• File Type:
  [PDF - 1.79 MB ]