Structural Basis for the Canonical and Non-canonical PAM Recognition by CRISPR-Cpf1
Name
nihms966209.pdf
Description
Accepted version
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1.72 MB
Format
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Checksum (MD5)
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Author(s)
Yamano, Takashi
Zetsche, Bernd
Ishitani, Ryuichiro
Zhang, Feng
Nishimasu, Hiroshi
Nureki, Osamu
Date Issued
August 2017
Journal
Molecular Cell
Publisher
Elsevier BV
Citation
Yamano, Takashi et al. "Structural Basis for the Canonical and Non-canonical PAM Recognition by CRISPR-Cpf1." Molecular Cell 67, 4 (August 2017): P633-645.e3 © 2017 Elsevier Inc.
Version
Author's final manuscript
Abstract
The RNA-guided Cpf1 (also known as Cas12a) nuclease associates with a CRISPR RNA (crRNA) and cleaves the double-stranded DNA target complementary to the crRNA guide. The two Cpf1 orthologs from Acidaminococcus sp. (AsCpf1) and Lachnospiraceae bacterium (LbCpf1) have been harnessed for eukaryotic genome editing. Cpf1 requires a specific nucleotide sequence, called a protospacer adjacent motif (PAM), for target recognition. Besides the canonical TTTV PAM, Cpf1 recognizes suboptimal C-containing PAMs. Here, we report four crystal structures of LbCpf1 in complex with the crRNA and its target DNA containing either TTTA, TCTA, TCCA, or CCCA as the PAM. These structures revealed that, depending on the PAM sequences, LbCpf1 undergoes conformational changes to form altered interactions with the PAM-containing DNA duplexes, thereby achieving the relaxed PAM recognition. Collectively, the present structures advance our mechanistic understanding of the PAM-dependent, crRNA-guided DNA cleavage by the Cpf1 family nucleases. Keywords: CRISPR-Cas; Cas12a; Cpf1; crystal structure; protospacer adjacent motif
MIT Department
Broad Institute of MIT and Harvard
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/j.molcel.2017.06.035
