8PM4

Cryo-EM structure of the Cas12m-crRNA-target DNA complex

8PM4 の概要

• エントリーDOI: 10.2210/pdb8pm4/pdb
• EMDBエントリー: 17757
• 分子名称: Transposase, crRNA, chain B, DNA oligoduplex, target strand, chain C, ... (4 entities in total)
• 機能のキーワード: cas12, cas12m, crispr-cas, ruvc, crrna, pam, rna binding protein
• 由来する生物種: Gordonia otitidis NBRC 100426; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 114075.30

構造登録者

Sasnauskas, G.,Tamulaitiene, G.,Karvelis, T.,Bigelyte, G.,Siksnys, V. (登録日: 2023-06-28, 公開日: 2024-02-07, 最終更新日: 2024-11-13)

主引用文献

Bigelyte, G.,Duchovska, B.,Zedaveinyte, R.,Sasnauskas, G.,Sinkunas, T.,Dalgediene, I.,Tamulaitiene, G.,Silanskas, A.,Kazlauskas, D.,Valancauskas, L.,Madariaga-Marcos, J.,Seidel, R.,Siksnys, V.,Karvelis, T.
Innate programmable DNA binding by CRISPR-Cas12m effectors enable efficient base editing.
Nucleic Acids Res., 52:3234-3248, 2024

PubMed Abstract: Cas9 and Cas12 nucleases of class 2 CRISPR-Cas systems provide immunity in prokaryotes through RNA-guided cleavage of foreign DNA. Here we characterize a set of compact CRISPR-Cas12m (subtype V-M) effector proteins and show that they provide protection against bacteriophages and plasmids through the targeted DNA binding rather than DNA cleavage. Biochemical assays suggest that Cas12m effectors can act as roadblocks inhibiting DNA transcription and/or replication, thereby triggering interference against invaders. Cryo-EM structure of Gordonia otitidis (Go) Cas12m ternary complex provided here reveals the structural mechanism of DNA binding ensuring interference. Harnessing GoCas12m innate ability to bind DNA target we fused it with adenine deaminase TadA-8e and showed an efficient A-to-G editing in Escherichia coli and human cells. Overall, this study expands our understanding of the functionally diverse Cas12 protein family, revealing DNA-binding dependent interference mechanism of Cas12m effectors that could be harnessed for engineering of compact base-editing tools.

PubMed: 38261981
DOI: 10.1093/nar/gkae016

実験手法

ELECTRON MICROSCOPY (2.93 Å)