+検索条件
-Structure paper
タイトル | Structure of the miniature type V-F CRISPR-Cas effector enzyme. |
---|---|
ジャーナル・号・ページ | Mol Cell, Vol. 81, Issue 3, Page 558-570.e3, Year 2021 |
掲載日 | 2021年2月4日 |
![]() | Satoru N Takeda / Ryoya Nakagawa / Sae Okazaki / Hisato Hirano / Kan Kobayashi / Tsukasa Kusakizako / Tomohiro Nishizawa / Keitaro Yamashita / Hiroshi Nishimasu / Osamu Nureki / ![]() |
PubMed 要旨 | RNA-guided DNA endonucleases derived from CRISPR-Cas adaptive immune systems are widely used as powerful genome-engineering tools. Among the diverse CRISPR-Cas nucleases, the type V-F Cas12f (also ...RNA-guided DNA endonucleases derived from CRISPR-Cas adaptive immune systems are widely used as powerful genome-engineering tools. Among the diverse CRISPR-Cas nucleases, the type V-F Cas12f (also known as Cas14) proteins are exceptionally compact and associate with a guide RNA to cleave single- and double-stranded DNA targets. Here, we report the cryo-electron microscopy structure of Cas12f1 (also known as Cas14a) in complex with a guide RNA and its target DNA. Unexpectedly, the structure revealed that two Cas12f1 molecules assemble with the single guide RNA to recognize the double-stranded DNA target. Each Cas12f1 protomer adopts a different conformation and plays distinct roles in nucleic acid recognition and DNA cleavage, thereby explaining how the miniature Cas12f1 enzyme achieves RNA-guided DNA cleavage as an "asymmetric homodimer." Our findings augment the mechanistic understanding of diverse CRISPR-Cas nucleases and provide a framework for the development of compact genome-engineering tools critical for therapeutic genome editing. |
![]() | ![]() ![]() |
手法 | EM (単粒子) |
解像度 | 3.3 Å |
構造データ | EMDB-30299, PDB-7c7l: |
化合物 | ![]() ChemComp-ZN: |
由来 |
|
![]() | RNA BINDING PROTEIN/RNA/DNA / Cas12f / Cas14 / sgRNA / target DNA / CRISPR / RNA BINDING PROTEIN-RNA-DNA complex |