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

Cell. 2015 Oct 22;163(3):759-71. doi: 10.1016/j.cell.2015.09.038. Epub 2015 Sep 25.

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 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • CRISPR-Cas Systems*
  • Endonucleases / chemistry
  • Endonucleases / genetics*
  • Francisella / enzymology
  • Francisella / genetics*
  • Genetic Engineering / methods*
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • RNA, Guide / genetics
  • Sequence Alignment

Substances

  • RNA, Guide
  • Endonucleases