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

   

Authors:	Gao, P.; Yang, H.; Rajashankar, K. R.; Huang, Z. W.; Patel, D. J.
Article Title:	Type V CRISPR-Cas Cpf1 endonuclease employs a unique mechanism for crRNA-mediated target DNA recognition
Abstract:	CRISPR-Cas9 and CRISPR-Cpf1 systems have been successfully harnessed for genome editing. In the CRIS-PR-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.
Keywords:	bacterial immunity; system; crystal-structure; escherichia-coli; cleavage; defense; prokaryotes; seed sequence; genome editing; crispr-cas; cpf1; crrna; guided surveillance complex; dual-rna
Journal Title:	Cell Research
Volume:	26
Issue:	8
ISSN:	1001-0602
Publisher:	Nature Publishing Group
Date Published:	2016-08-01
Start Page:	901
End Page:	913
Language:	English
ACCESSION:	WOS:000380923900008
DOI:	10.1038/cr.2016.88
PROVIDER:	wos
PMCID:	PMC4973337
PUBMED:	27444870
Notes:	Article -- Source: Wos

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