2019年2月7日 · We show that AcrIIA2 binds SpyCas9 at a position similar to the target DNA binding region. More specifically, AcrIIA2 interacts with the protospacer adjacent motif (PAM) recognition residues of Cas9, preventing target double-stranded DNA (dsDNA) detection.

2017年4月27日 · Our data show that AcrIIA2 and AcrIIA4 interact with SpyCas9 in a sgRNA-dependent manner. The structure reveals that AcrIIA4 inhibits SpyCas9 activity by structurally mimicking the PAM to occupy...

The expression of AcrIIA4 yields robust inhibition of Cas9-based phage targeting at all induction levels of Cas9, while AcrIIA2 provides limited activity under the strongest Cas9 induction conditions but functioned well at lower levels of Cas9 (Figure 4C).

2019年2月7日 · We show here that AcrIIA2 and its more potent homolog, AcrIIA2b, prevent Cas9 binding to DNA by occluding protein residues required for DNA binding. Cryo-EM-determined structures of AcrIIA2 or AcrIIA2b bound to S. pyogenes Cas9 reveal a mode of competitive inhibition of DNA binding that is distinct from other known Acrs.

We show here that AcrIIA2 and its more potent homolog, AcrIIA2b, prevent Cas9 binding to DNA by occluding protein residues required for DNA binding. Cryo-EM-determined structures of AcrIIA2 or AcrIIA2b bound to S. pyogenes Cas9 reveal a mode of competitive inhibition of DNA binding that is distinct from other known Acrs.

2023年4月1日 · Several anti-CRISPRs have been shown to use DNA mimicry to prevent target DNA binding by Cas9. For example, AcrIIA2 and AcrIIA4 were found to bind much more strongly to the Cas9 RNP compared to apo-Cas9 and also were unable to inhibit Cas9 activity when added after target DNA, suggesting competitive binding. 37, 38, 39, 40 Structures of each ...

AcrIIC2 inhibits crRNA loading into Cas9 preventing proper complex assembly. AcrIIA2, AcrIIA4, AcrIIC3, AcrIIC4, and AcrIIC5 prevent the complex from recognizing target DNA. Following target recognition, Cas9 creates a double stranded DNA break target, leading to its destruction.

2024年12月1日 · In this study, through microsecond atomistic molecular dynamics simulations, we demonstrated the remarkable flexibility of Streptococcus pyogenes Cas9 (SpyCas9) and its conformational adaptability during interactions with AcrIIA4 and AcrIIA2. Specifically, we demonstrated that the binding of AcrIIA4 and AcrIIA2 to SpyCas9 induces a ...

冷冻电镜测定的 AcrIIA2 或 AcrIIA2b 与化脓性链球菌 Cas9 结合的结构揭示了一种与其他已知 Acrs 不同的 DNA 结合竞争性抑制模式。 AcrIIA2 和 AcrIIA2b 对 Cas9 抑制的温度依赖性差异源于 Cas9 上抑制剂结构和局部抑制剂结合环境的差异。

2019年6月26日 · We demonstrate that AcrIIC2 Nme inhibits Cas9 through interactions with the positively charged bridge helix, thereby preventing sgRNA loading. In vivo phage plaque assays and in vitro DNA...