Mechanism of Cas9 inhibition by AcrIIA11 DOI Creative Commons

Kaylee E. Dillard,

Hongshan Zhang,

Lianne Z Dubbs

et al.

Nucleic Acids Research, Journal Year: 2025, Volume and Issue: 53(8)

Published: April 22, 2025

Abstract Mobile genetic elements evade CRISPR–Cas adaptive immunity by encoding anti-CRISPR proteins (Acrs). Acrs inactivate systems via diverse mechanisms but generally coevolve with a narrow subset of Cas effectors that share high sequence similarity. Here, we demonstrate AcrIIA11 inhibits Streptococcus pyogenes (Sp), Staphylococcus aureus (Sa), and Francisella novicida (Fn) Cas9s in vitro human cells. Single-molecule imaging reveals hinders SaCas9 target search reducing its diffusion on nonspecific DNA. DNA cleavage is inhibited because the AcrIIA11:SaCas9 complex binds to protospacer adjacent motif (PAM)-rich off-target sites, preventing from reaching target. also greatly slows down after reaches site. A negative-stain electron microscopy reconstruction an RNP heterodimer assembles 1:1 stoichiometry. Physical AcrIIA11–Cas9 interactions across type IIA IIB correlate nuclease inhibition support broad-spectrum activity. These results add kinetic mechanism phage-CRISPR arms race.

Language: Английский

Recommendations for robust and reproducible research on ferroptosis DOI
Eikan Mishima, Toshitaka Nakamura, Sebastian Doll

et al.

Nature Reviews Molecular Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: April 9, 2025

Language: Английский

Citations

0
Mechanism of Cas9 inhibition by AcrIIA11 DOI Creative Commons

Kaylee E. Dillard,

Hongshan Zhang,

Lianne Z Dubbs

et al.

Nucleic Acids Research, Journal Year: 2025, Volume and Issue: 53(8)

Published: April 22, 2025

Abstract Mobile genetic elements evade CRISPR–Cas adaptive immunity by encoding anti-CRISPR proteins (Acrs). Acrs inactivate systems via diverse mechanisms but generally coevolve with a narrow subset of Cas effectors that share high sequence similarity. Here, we demonstrate AcrIIA11 inhibits Streptococcus pyogenes (Sp), Staphylococcus aureus (Sa), and Francisella novicida (Fn) Cas9s in vitro human cells. Single-molecule imaging reveals hinders SaCas9 target search reducing its diffusion on nonspecific DNA. DNA cleavage is inhibited because the AcrIIA11:SaCas9 complex binds to protospacer adjacent motif (PAM)-rich off-target sites, preventing from reaching target. also greatly slows down after reaches site. A negative-stain electron microscopy reconstruction an RNP heterodimer assembles 1:1 stoichiometry. Physical AcrIIA11–Cas9 interactions across type IIA IIB correlate nuclease inhibition support broad-spectrum activity. These results add kinetic mechanism phage-CRISPR arms race.

Language: Английский

Citations

0