7VTN

Cryo-EM structure of the Cas13bt3-crRNA-target RNA ternary complex

7VTN の概要

• エントリーDOI: 10.2210/pdb7vtn/pdb
• EMDBエントリー: 32118
• 分子名称: Cas13bt3, crRNA, target RNA (3 entities in total)
• 機能のキーワード: crispr-cas, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: Planctomycetes bacterium; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 118087.18

構造登録者

Nakagawa, R.,Soumya, K.,Han, A.,Takeda, N.S.,Tomita, A.,Hirano, H.,Kusakizako, T.,Tomohiro, N.,Yamashita, K.,Feng, Z.,Nishimasu, H.,Nureki, O. (登録日: 2021-10-30, 公開日: 2022-09-07, 最終更新日: 2024-06-26)

主引用文献

Nakagawa, R.,Kannan, S.,Altae-Tran, H.,Takeda, S.N.,Tomita, A.,Hirano, H.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Zhang, F.,Nishimasu, H.,Nureki, O.
Structure and engineering of the minimal type VI CRISPR-Cas13bt3.
Mol.Cell, 82:3178-3192.e5, 2022

PubMed Abstract: Type VI CRISPR-Cas13 effector enzymes catalyze RNA-guided RNA cleavage and have been harnessed for various technologies, such as RNA detection, targeting, and editing. Recent studies identified Cas13bt3 (also known as Cas13X.1) as a miniature Cas13 enzyme, which can be used for knockdown and editing of target transcripts in mammalian cells. However, the action mechanism of the compact Cas13bt3 remains unknown. Here, we report the structures of the Cas13bt3-guide RNA complex and the Cas13bt3-guide RNA-target RNA complex. The structures revealed how Cas13bt3 recognizes the guide RNA and its target RNA and provided insights into the activation mechanism of Cas13bt3, which is distinct from those of the other Cas13a/d enzymes. Furthermore, we rationally engineered enhanced Cas13bt3 variants and ultracompact RNA base editors. Overall, this study improves our mechanistic understanding of the CRISPR-Cas13 enzymes and paves the way for the development of efficient Cas13-mediated transcriptome modulation technologies.

PubMed: 36027912
DOI: 10.1016/j.molcel.2022.08.001

実験手法

ELECTRON MICROSCOPY (3.38 Å)