PAM-dependent target DNA recognition and cleavage by C2c1 CRISPR-Cas endonuclease Journal Article
| Authors: | Yang, H.; Gao, P.; Rajashankar, K. R.; Patel, D. J. |
| Article Title: | PAM-dependent target DNA recognition and cleavage by C2c1 CRISPR-Cas endonuclease |
| Abstract: | 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 Alicyclobacillus 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. © 2016 Elsevier Inc. |
| Keywords: | structure; binary complex with sgrna; c2c1; genome editing tool; ruvc catalytic pocket; sequence-specific pam recognition; ternary complex with added dna; type v crispr-cas endonuclease |
| Journal Title: | Cell |
| Volume: | 167 |
| Issue: | 7 |
| ISSN: | 0092-8674 |
| Publisher: | Cell Press |
| Date Published: | 2016-12-15 |
| Start Page: | 1814 |
| End Page: | 1828.e12 |
| Language: | English |
| DOI: | 10.1016/j.cell.2016.11.053 |
| PROVIDER: | scopus |
| PUBMED: | 27984729 |
| PMCID: | PMC5278635 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 3 January 2017 -- Source: Scopus |
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