Proteasomal control of anti-CRISPRs for the regulation of CRISPR/Cas9 activity using Cas9-ACROBAT
Timothy D. MartiniD, Emma V. WatsoniD, Mei Yuk Choi, Behnam NabetiD, Nathanael S. GrayiD, Qikai XuiD, Stephen Joseph ElledgeiD
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.
02 /Specification & Classification
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