Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs

Kale Kundert (), James E. Lucas, Kyle E. Watters, Christof Fellmann, Andrew H. Ng, Benjamin M. Heineike, Christina M. Fitzsimmons, Benjamin L. Oakes, Jiuxin Qu, Neha Prasad, Oren S. Rosenberg, David F. Savage, Hana El-Samad, Jennifer A. Doudna and Tanja Kortemme ()
Additional contact information
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: University of California San Francisco
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: Chan Zuckerberg Biohub
Jennifer A. Doudna: University of California Berkeley
Tanja Kortemme: Chan Zuckerberg Biohub

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-019-09985-2

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