6W5C

Cryo-EM structure of Cas12i(E894A)-crRNA-dsDNA complex

6W5C の概要

• エントリーDOI: 10.2210/pdb6w5c/pdb
• EMDBエントリー: 21541 21551 21552
• 分子名称: Cas12i, TS, NTS, ... (5 entities in total)
• 機能のキーワード: crispr, hydrolase-dna-rna complex, hydrolase/dna/rna
• 由来する生物種: Lachnospiraceae bacterium ND2006; 詳細
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 161424.09

構造登録者

Chang, L.,Li, Z.,Zhang, H. (登録日: 2020-03-13, 公開日: 2020-09-16, 最終更新日: 2024-03-06)

主引用文献

Zhang, H.,Li, Z.,Xiao, R.,Chang, L.
Mechanisms for target recognition and cleavage by the Cas12i RNA-guided endonuclease.
Nat.Struct.Mol.Biol., 27:1069-1076, 2020

PubMed Abstract: Cas12i is a recently identified type V CRISPR-Cas endonuclease that predominantly cleaves the non-target strand of a double-stranded DNA substrate. This nicking activity of Cas12i could potentially be used for genome editing with high specificity. To elucidate its mechanisms for target recognition and cleavage, we determined cryo-EM structures of Cas12i in multiple functional states. Cas12i pre-orders a seven-nucleotide seed sequence of the crRNA for target recognition and undergoes a two-step activation through crRNA-DNA hybridization. Formation of 14 base pairs activates the nickase activity, and 28-bp hybridization promotes cleavage of the target strand. The atomic structures and mechanistic insights gained should facilitate the manipulation of Cas12i for genome editing applications.

PubMed: 32895556
DOI: 10.1038/s41594-020-0499-0

実験手法

ELECTRON MICROSCOPY (2.9 Å)