Precise Regulation of Cas9-Mediated Genome Engineering by Anti-CRISPR-Based Inducible CRISPR Controllers

Dose-dependent control over Cas9 editing and dCas9 induction was achieved, significantly improving the specificity and biosafety of the CRISPR/Cas9 system.

• A synthetic controller using a chemically inducible anti-CRISPR protein was developed to regulate Cas9 activity.
• The anti-CRISPR protein AcrIIA4 can inhibit Cas9 DNA binding when trimethoprim is added.
• Reduced off-target activity was observed in mammalian cells with the engineered system.
• An inducible promoter system, Acr-OFF, was created to provide additional control over CRISPR activity.
• The system is regulated by trimethoprim, which is approved by the FDA.

AI simplified