期刊
MOLECULAR CELL
卷 65, 期 2, 页码 310-322出版社
CELL PRESS
DOI: 10.1016/j.molcel.2016.11.040
关键词
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资金
- Natural Science Foundation of China [91440201, 31630015, 31571335, 31400640]
- Chinese Ministry of Science and Technology [2014CB910102]
- Chinese Academy of Sciences [XDB08010203]
C2c1 is a type V-B CRISPR-Cas system dual-RNAguided DNA endonuclease. Here, we report the crystal structure of Alicyclobacillus acidoterrestris C2c1 in complex with a chimeric single-molecule guide RNA ( sgRNA). AacC2c1 exhibits a bi-lobed architecture consisting of a REC and NUC lobe. The sgRNA scaffold forms a tetra-helical structure, distinct from previous predictions. The crRNA is located in the central channel of C2c1, and the tracrRNA resides in an external surface groove. Although AacC2c1 lacks a PAM-interacting domain, our analysis revealed that the PAM duplex has a similar binding position found in Cpf1. Importantly, C2c1-sgRNA system is highly sensitive to singlenucleotide mismatches between guide RNA and target DNA. The resulting reduction in off-target cleavage renders C2c1 a valuable addition to the current arsenal of genome-editing tools. Together, our findings indicate that sgRNA assembly is achieved through a mechanism distinct from that reported previously for Cas9 or Cpf1 endonucleases.
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