期刊
CELL
卷 167, 期 7, 页码 1814-+出版社
CELL PRESS
DOI: 10.1016/j.cell.2016.11.053
关键词
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资金
- NIGMS [P41 GM103403]
- U.S. Department of Energy [DE-AC02-06CH11357]
- NIHORIP HEI grant [S10 RR029205]
- NIH [GM104962]
- Memorial Sloan Kettering Cancer Center Core Grant [P30CA008748]
- Cancer Research Institute Irvington Postdoctoral Fellowship
- Institute of Biophysics, Beijing, China
C2c1 is a newly identified guide RNA-mediated type V-B CRISPR-Cas endonuclease that site-specifically targets and cleaves both strands of target DNA. We have determined crystal structures of Alicyclo-bacillus acidoterrestris C2c1 (AacC2c1) bound to sgRNA as a binary complex and to target DNAs as ternary complexes, thereby capturing catalytically competent conformations of AacC2c1 with both target and non-target DNA strands independently positioned within a single RuvC catalytic pocket. Moreover, C2c1-mediated cleavage results in a staggered seven-nucleotide break of target DNA. crRNA adopts a pre-ordered five-nucleotide A-form seed sequence in the binary complex, with release of an inserted tryptophan, facilitating zippering up of 20-bp guide RNA: target DNA heteroduplex on ternary complex formation. Notably, the PAM-interacting cleft adopts a locked'' conformation on ternary complex formation. Structural comparison of C2c1 ternary complexes with their Cas9 and Cpf1 counterparts highlights the diverse mechanisms adopted by these distinct CRISPR-Cas systems, thereby broadening and enhancing their applicability as genome editing tools.
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