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-Structure paper
Title | Structure of the type V-C CRISPR-Cas effector enzyme. |
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Journal, issue, pages | Mol Cell, Vol. 82, Issue 10, Page 1865-11877.e4, Year 2022 |
Publish date | May 19, 2022 |
Authors | Nina Kurihara / Ryoya Nakagawa / Hisato Hirano / Sae Okazaki / Atsuhiro Tomita / Kan Kobayashi / Tsukasa Kusakizako / Tomohiro Nishizawa / Keitaro Yamashita / David A Scott / Hiroshi Nishimasu / Osamu Nureki / |
PubMed Abstract | RNA-guided CRISPR-Cas nucleases are widely used as versatile genome-engineering tools. Recent studies identified functionally divergent type V Cas12 family enzymes. Among them, Cas12c2 binds a CRISPR ...RNA-guided CRISPR-Cas nucleases are widely used as versatile genome-engineering tools. Recent studies identified functionally divergent type V Cas12 family enzymes. Among them, Cas12c2 binds a CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA) and recognizes double-stranded DNA targets with a short TN PAM. Here, we report the cryo-electron microscopy structures of the Cas12c2-guide RNA binary complex and the Cas12c2-guide RNA-target DNA ternary complex. The structures revealed that the crRNA and tracrRNA form an unexpected X-junction architecture, and that Cas12c2 recognizes a single T nucleotide in the PAM through specific hydrogen-bonding interactions with two arginine residues. Furthermore, our biochemical analyses indicated that Cas12c2 processes its precursor crRNA to a mature crRNA using the RuvC catalytic site through a unique mechanism. Collectively, our findings improve the mechanistic understanding of diverse type V CRISPR-Cas effectors. |
External links | Mol Cell / PubMed:35366394 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.7 - 3.0 Å |
Structure data | EMDB-31807, PDB-7v93: EMDB-31808, PDB-7v94: |
Source |
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Keywords | RNA BINDING PROTEIN/RNA / cas12c / c2c3 / sgRNA / CRISPR / RNA binding protein-RNA complex / RNA BINDING PROTEIN/RNA/DNA / target DNA / RNA binding protein-RNA-DNA complex |