Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs
Kale Kundert(University of California, San Francisco), James E. Lucas(University of California, San Francisco), Kyle E. Watters(University of California, Berkeley), Christof Fellmann(University of California, Berkeley), Andrew H. Ng(University of California, San Francisco), Benjamin M. Heineike(University of California, San Francisco), Christina M. Fitzsimmons(National Institutes of Health), Benjamin L. Oakes(University of California, Berkeley), Jiuxin Qu(University of California, San Francisco), Neha Prasad(University of California, San Francisco), Oren S. Rosenberg(University of California, San Francisco), David F. Savage(University of California, Berkeley), Hana El‐Samad(University of California, San Francisco), Jennifer A. Doudna(Howard Hughes Medical Institute), Tanja Kortemme(University of California, San Francisco)
Cited by 191Open Access
Abstract
The CRISPR-Cas9 system provides the ability to edit, repress, activate, or mark any gene (or DNA element) by pairing of a programmable single guide RNA (sgRNA) with a complementary sequence on the DNA target. Here we present a new method for small-molecule control of CRISPR-Cas9 function through insertion of RNA aptamers into the sgRNA. We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands. Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.
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