The compact Casπ (Cas12l) 'bracelet' provides a unique structural platform for DNA manipulation.
Cell Res
; 33(3): 229-244, 2023 Mar.
Article
em En
| MEDLINE
| ID: mdl-36650285
ABSTRACT
CRISPR-Cas modules serve as the adaptive nucleic acid immune systems for prokaryotes, and provide versatile tools for nucleic acid manipulation in various organisms. Here, we discovered a new miniature type V system, CRISPR-Casπ (Cas12l) (~860 aa), from the environmental metagenome. Complexed with a large guide RNA (~170 nt) comprising the tracrRNA and crRNA, Casπ (Cas12l) recognizes a unique 5' C-rich PAM for DNA cleavage under a broad range of biochemical conditions, and generates gene editing in mammalian cells. Cryo-EM study reveals a 'bracelet' architecture of Casπ effector encircling the DNA target at 3.4 Å resolution, substantially different from the canonical 'two-lobe' architectures of Cas12 and Cas9 nucleases. The large guide RNA serves as a 'two-arm' scaffold for effector assembly. Our study expands the knowledge of DNA targeting mechanisms by CRISPR effectors, and offers an efficient but compact platform for DNA manipulation.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
DNA
/
Edição de Genes
Idioma:
En
Revista:
Cell Res
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
China