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Cell Res. 2016 Aug;26(8):901-13. doi: 10.1038/cr.2016.88. Epub 2016 Jul 22.

Type V CRISPR-Cas Cpf1 endonuclease employs a unique mechanism for crRNA-mediated target DNA recognition.

Author information

1
Key Laboratory of Infection and Immunity, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
2
Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA.
3
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA.
4
NE-CAT, Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60349, USA.
5
School of Life Science and Technology, Harbin Institute of Technology, Harbin 150880, China.

Abstract

CRISPR-Cas9 and CRISPR-Cpf1 systems have been successfully harnessed for genome editing. In the CRISPR-Cas9 system, the preordered A-form RNA seed sequence and preformed protein PAM-interacting cleft are essential for Cas9 to form a DNA recognition-competent structure. Whether the CRISPR-Cpf1 system employs a similar mechanism for target DNA recognition remains unclear. Here, we have determined the crystal structure of Acidaminococcus sp. Cpf1 (AsCpf1) in complex with crRNA and target DNA. Structural comparison between the AsCpf1-crRNA-DNA ternary complex and the recently reported Lachnospiraceae bacterium Cpf1 (LbCpf1)-crRNA binary complex identifies a unique mechanism employed by Cpf1 for target recognition. The seed sequence required for initial DNA interrogation is disordered in the Cpf1-cRNA binary complex, but becomes ordered upon ternary complex formation. Further, the PAM interacting cleft of Cpf1 undergoes an "open-to-closed" conformational change upon target DNA binding, which in turn induces structural changes within Cpf1 to accommodate the ordered A-form seed RNA segment. This unique mechanism of target recognition by Cpf1 is distinct from that reported previously for Cas9.

PMID:
27444870
PMCID:
PMC4973337
DOI:
10.1038/cr.2016.88
[Indexed for MEDLINE]
Free PMC Article

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