7WAH

Structure of Cas7-11 in complex with guide RNA and target RNA

Summary for 7WAH

• Entry DOI: 10.2210/pdb7wah/pdb
• EMDB information: 32385
• Descriptor: crRNA (39-MER), tgRNA (5'-R(P*UP*AP*CP*CP*CP*AP*UP*GP*UP*CP*GP*AP*AP*GP*AP*CP*AP*AP*CP*AP*AP*AP*G)-3'), CRISPR-associated RAMP family protein, ... (4 entities in total)
• Functional Keywords: crispr, rnase, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Escherichia coli; More
• Total number of polymer chains: 3
• Total formula weight: 205751.40

Authors

Kato, K.,Okazaki, S.,Isayama, Y.,Nishizawa, T.,Nishimasu, H. (deposition date: 2021-12-14, release date: 2022-06-15, Last modification date: 2024-06-26)

Primary citation

Kato, K.,Zhou, W.,Okazaki, S.,Isayama, Y.,Nishizawa, T.,Gootenberg, J.S.,Abudayyeh, O.O.,Nishimasu, H.
Structure and engineering of the type III-E CRISPR-Cas7-11 effector complex.
Cell, 185:2324-2337.e16, 2022

PubMed Abstract: The type III-E CRISPR-Cas effector Cas7-11, with dual RNase activities for precursor CRISPR RNA (pre-crRNA) processing and crRNA-guided target RNA cleavage, is a new platform for bacterial and mammalian RNA targeting. We report the 2.5-Å resolution cryoelectron microscopy structure of Cas7-11 in complex with a crRNA and its target RNA. Cas7-11 adopts a modular architecture comprising seven domains (Cas7.1-Cas7.4, Cas11, INS, and CTE) and four interdomain linkers. The crRNA 5' tag is recognized and processed by Cas7.1, whereas the crRNA spacer hybridizes with the target RNA. Consistent with our biochemical data, the catalytic residues for programmable cleavage in Cas7.2 and Cas7.3 neighbor the scissile phosphates before the flipped-out fourth and tenth nucleotides in the target RNA, respectively. Using structural insights, we rationally engineered a compact Cas7-11 variant (Cas7-11S) for single-vector AAV packaging for transcript knockdown in human cells, enabling in vivo Cas7-11 applications.

PubMed: 35643083
DOI: 10.1016/j.cell.2022.05.003

Experimental method

ELECTRON MICROSCOPY (2.45 Å)