9IZR

Cryo-EM structure of CasLambda2-crRNA-target DNA ternary complex in the NTS-cleaving state

9IZR の概要

• エントリーDOI: 10.2210/pdb9izr/pdb
• EMDBエントリー: 61040
• 分子名称: Cas Lambda2, RNA (58-MER), DNA (40-MER), ... (8 entities in total)
• 機能のキーワード: crispr-cas12, genome engineering, hydrolase-rna-dna complex, dna binding protein
• 由来する生物種: unidentified; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 134671.01

構造登録者

Omura, S.N.,Hirano, H.,Itoh, Y.,Nureki, O. (登録日: 2024-08-01, 公開日: 2025-06-11, 最終更新日: 2025-06-25)

主引用文献

Omura, S.N.,Alfonse, L.E.,Ornstein, A.,Morinaga, H.,Hirano, H.,Itoh, Y.,Munoz, G.,Garrity, A.J.,Hoffman, G.R.,DiTommaso, T.,Yan, W.X.,Cheng, D.R.,Scott, D.A.,Maben, Z.,Nureki, O.
Structural basis for target DNA cleavage and guide RNA processing by CRISPR-Cas lambda 2.
Commun Biol, 8:876-876, 2025

PubMed Abstract: RNA-guided CRISPR-Cas nucleases are widely used as versatile genome-engineering tools. Among the diverse CRISPR-Cas effectors, CRISPR-Casλ-also referred to as Cas12n-is a recently identified miniature type V nuclease encoded in phage genomes. Given its demonstrated nuclease activity in both mammalian and plant cells, Casλ has emerged as a promising candidate for genome-editing applications. However, the precise molecular mechanisms of Casλ family enzymes remain poorly understood. In this study, we report the identification and detailed biochemical and structural characterizations of CRISPR-Casλ2. The cryo-electron microscopy structures of Casλ2 in five different functional states unveiled the dynamic domain rearrangements during its activation. Our biochemical analyses indicated that Casλ2 processes its precursor crRNA to a mature crRNA using the RuvC active site through a unique ruler mechanism, in which Casλ2 defines the spacer length of the mature crRNA. Furthermore, structural comparisons of Casλ2 with Casλ1 and CasΦ highlighted the diversity and conservation of phage-encoded type V CRISPR-Cas enzymes. Collectively, our findings augment the mechanistic understanding of diverse CRISPR-Cas nucleases and establish a framework for rational engineering of the CRISPR-Casλ-based genome-editing platform.

PubMed: 40473912
DOI: 10.1038/s42003-025-08300-8

実験手法

ELECTRON MICROSCOPY (2.93 Å)