2018年10月30日 · We engineered optogenetic anti-CRISPR variants comprising hybrids of AcrIIA4, a potent Streptococcus pyogenes Cas9 inhibitor, and the LOV2 photosensor from Avena sativa. Coexpression of these...

2023年7月1日 · More recently, a separate group showed that circularly permuted LOV2 (cpLOV2, which can provide distinct “caging surfaces” relative to the non-permuted domain) can also control anti-CRISPR functions, 39 affording additional options for effective fusions to the ever-growing roster of Acr proteins.

通过 诱导型启动子 、光或小分子调控Acr蛋白，可以实现CRISPR-Cas活性的快速、动态和空间控制。例如，将AcrA4II融合到光敏LOV2结构域，使得抗CRISPR活性可以被蓝光控制(Bubeck等,2018)。 降低CRISPR-Cas编辑在组织中的细胞毒性

2023年11月1日 · We combined Acr with the light-oxygen-voltage 2 (LOV2) photosensor from Avena sativa according to previous studies (Supplementary Figure S4A) (29, 30). We also optimized the PICI system to simplify the previous procedures by redesigning the complete plasmid for expressing Cas9 and Acr using araBAD and …

2020年12月16日 · Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein. Building on our previous Acr engineering work, we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa.

2020年4月13日 · Anti-CRISPR (Acr) proteins are powerful tools to control CRISPR–Cas technologies. However, the available Acr repertoire is limited to naturally occurring variants. Here, we applied...

We engineered optogenetic anti-CRISPR variants comprising hybrids of AcrIIA4, a potent Streptococcus pyogenes Cas9 inhibitor, and the LOV2 photosensor from Avena sativa. Coexpression of these proteins with CRISPR-Cas9 effectors enabled light-mediated genome and epigenome editing, and revealed rapid Cas9 genome targeting in human cells.

2020年10月13日 · CASANOVA, which stands for ‘CRISPR-Cas9 activity switching via a novel optogenetic variant of AcrIIA4’, represents a chimeric protein made of AcrIIA4 fused to the LOV2 photosensor domain from the Avena sativa phototropin-1 protein.

2023年12月11日 · Anti-CRISPR (Acr) proteins are encoded by mobile genetic elements to overcome the CRISPR immunity of prokaryotes, displaying promises as controllable tools for modulating CRISPR-based applications. However, characterizing novel anti-CRISPR proteins and exploiting Acr-related technologies is a rather …

2024年3月6日 · The resulting Acr-LOV2 fusion protein, named CASANOVA (for CRISPR-Cas9 activity switching via a novel optogenetic variant of AcrIIA4), blocks SpyCas9 activity in the dark similarly to wild-type AcrIIA4. However, in the presence of blue light, the LOV2 photosensory domain gains flexibility by undocking and unfolding of its terminal helices.