6DTD

High-resolution crystal structure of Cas13b from Prevotella buccae

6DTD の概要

• エントリーDOI: 10.2210/pdb6dtd/pdb
• 分子名称: nuclease, RNA (37-MER), CITRIC ACID, ... (7 entities in total)
• 機能のキーワード: nuclease, hydrolase-rna complex, hydrolase/rna
• 由来する生物種: Prevotella buccae; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 148288.56

構造登録者

Slaymaker, I.M.S.,Zhang, F.Z. (登録日: 2018-06-15, 公開日: 2019-02-20, 最終更新日: 2024-03-13)

主引用文献

Slaymaker, I.M.,Mesa, P.,Kellner, M.J.,Kannan, S.,Brignole, E.,Koob, J.,Feliciano, P.R.,Stella, S.,Abudayyeh, O.O.,Gootenberg, J.S.,Strecker, J.,Montoya, G.,Zhang, F.
High-Resolution Structure of Cas13b and Biochemical Characterization of RNA Targeting and Cleavage.
Cell Rep, 26:3741-3751.e5, 2019

PubMed Abstract: Type VI CRISPR-Cas systems contain programmable single-effector RNA-guided RNases, including Cas13b, one of the four known family members. Cas13b, which has been used for both RNA editing and nucleic acid detection, is unique among type VI CRISPR effectors in its linear domain architecture and CRISPR RNA (crRNA) structure. Here, we report the crystal structure of Prevotella buccae Cas13b (PbuCas13b) bound to crRNA at 1.65 Å resolution. This structure, combined with biochemical experiments assaying the stability, kinetics, and function of Cas13b, provides a mechanistic model for Cas13b target RNA recognition and identifies features responsible for target and cleavage specificity. Based on these observations, we generated Cas13b variants with altered cleavage preferences, which may expand the utility of nuclease-based RNA detection assays and other applications of Cas13b in mammalian cells.

PubMed: 30917325
DOI: 10.1016/j.celrep.2019.02.094

実験手法

X-RAY DIFFRACTION (1.65 Å)