7WAH

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

7WAH の概要

• エントリーDOI: 10.2210/pdb7wah/pdb
• EMDBエントリー: 32385
• 分子名称: 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)
• 機能のキーワード: crispr, rnase, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 205751.40

構造登録者

Kato, K.,Okazaki, S.,Isayama, Y.,Nishizawa, T.,Nishimasu, H. (登録日: 2021-12-14, 公開日: 2022-06-15, 最終更新日: 2025-06-18)

主引用文献

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

実験手法

ELECTRON MICROSCOPY (2.45 Å)