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

• DOI: 10.1021/acssynbio.0c00548
• Publication Date: 2021-05-19T06:16:52Z
• Authors: Surbhi Jain; Guanhua Xun; Shireen Abesteh; Sherri Ho; Manasi Lingamaneni; Teresa A. Martin; Ipek Tasan; Che Yang; Huimin Zhao
• Keywords: Genome Engineering; Trans-activating crRNA; Guide RNA

CRISPR/Cas9 is a powerful genome editing tool, but its off-target cleavage activity can result in unintended adverse outcomes for therapeutic applications. Here we report the design of simple tunable CRISPR controller which chemically inducible anti-CRISPR protein AcrIIA4 engineered to disable Cas9 DNA binding upon addition trimethoprim. Dose-dependent control over and dCas9 induction was achieved, drastically improved specificity biosafety system. We utilized as means interfere with activity. By fusing ligand-inducible destabilization domain DHFR(DD), show significantly reduced mammalian cells. Furthermore, describe new promoter system Acr-OFF based on controllers, regulated by an FDA-approved ligand