EMD-33983
Structure of CasPi with guide RNA and target DNA
EMD-33983
Single-particle3.36 Å
Deposition: 01/08/2022Map released: 15/02/2023
Last modified: 03/07/2024
Sample Organism:
Armatimonadetes bacterium
Sample: CasPi
Fitted models: 7yoj (Avg. Q-score: 0.361)
Deposition Authors: Li CP, Wang J
,
Liu JJ,
Zhang S,
Wang J ,
Liu JJ
Sample: CasPi
Fitted models: 7yoj (Avg. Q-score: 0.361)
Deposition Authors: Li CP, Wang J
,
Liu JJ,
Zhang S,
Wang J ,
Liu JJ
The compact Cas pi (Cas12l) 'bracelet' provides a unique structural platform for DNA manipulation.
Sun A,
Li CP,
Chen Z ,
Zhang S,
Li DY,
Yang Y,
Li LQ,
Zhao Y,
Wang K,
Li Z,
Liu J,
Liu S,
Wang J ,
Liu JG
(2023) Cell Res , 33 , 229 - 244
,
Zhang S,
Li DY,
Yang Y,
Li LQ,
Zhao Y,
Wang K,
Li Z,
Liu J,
Liu S,
Wang J ,
Liu JG
(2023) Cell Res , 33 , 229 - 244
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.
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.