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[Preprint]. 2024 May 13:2024.05.13.593596. [Version 1] doi: 10.1101/2024.05.13.593596

Proteasomal control of anti-CRISPRs for the regulation of CRISPR/Cas9 activity using Cas9-ACROBAT

Timothy D Martin, Emma V Watson, Mei Yuk Choi, Behnam Nabet, Nathanael S Gray, Qikai Xu, Stephen J Elledge
PMCID: PMC11118331  PMID: 38798327

ABSTRACT

Small molecule-mediated proteasomal degradation of proteins is a powerful tool for synthetic regulation of biological activity. To control Cas9 activity in cells, we engineered an anti-CRISPR protein, AcrIIA4, fused to a degradation (dTAG) or small molecule assisted shutoff (SMASh) tag. Co-expression of the tagged AcrIIA4 along with Cas9 and riboswitch-regulated sgRNAs enables precise tunable control of CRISPR activity by small molecule addition.

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