7N3P

Cryo-EM structure of the Cas12k-sgRNA-dsDNA complex

7N3P の概要

• エントリーDOI: 10.2210/pdb7n3p/pdb
• EMDBエントリー: 24143
• 分子名称: Cas12k, Single guide RNA, DNA (5'-D(*CP*AP*TP*GP*AP*CP*TP*TP*CP*TP*CP*AP*AP*CP*CP*GP*AP*GP*TP*TP*T)-3'), ... (4 entities in total)
• 機能のキーワード: crispr cas, rna binding protein, protein-rna-dna complex, rna binding protein-rna-dna complex, rna binding protein/rna/dna
• 由来する生物種: Scytonema hofmannii; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 174153.07

構造登録者

Chang, L.,Li, Z.,Xiao, R.,Wang, S.,Han, R. (登録日: 2021-06-01, 公開日: 2021-09-01, 最終更新日: 2025-06-04)

主引用文献

Xiao, R.,Wang, S.,Han, R.,Li, Z.,Gabel, C.,Mukherjee, I.A.,Chang, L.
Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
Mol.Cell, 81:4457-4466.e5, 2021

PubMed Abstract: The type V-K CRISPR-Cas system, featured by Cas12k effector with a naturally inactivated RuvC domain and associated with Tn7-like transposon for RNA-guided DNA transposition, is a promising tool for precise DNA insertion. To reveal the mechanism underlying target DNA recognition, we determined a cryoelectron microscopy (cryo-EM) structure of Cas12k from cyanobacteria Scytonema hofmanni in complex with a single guide RNA (sgRNA) and a double-stranded target DNA. Coupled with mutagenesis and in vitro DNA transposition assay, our results revealed mechanisms for the recognition of the GGTT protospacer adjacent motif (PAM) sequence and the structural elements of Cas12k critical for RNA-guided DNA transposition. These structural and mechanistic insights should aid in the development of type V-K CRISPR-transposon systems as tools for genome editing.

PubMed: 34450043
DOI: 10.1016/j.molcel.2021.07.043

実験手法

ELECTRON MICROSCOPY (3.65 Å)