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

Kundert, Kale; Lucas, James E; Watters, Kyle E; Fellmann, Christof; Ng, Andrew H; Heineike, Benjamin M; Fitzsimmons, Christina M; Oakes, Benjamin L et al. · Nat Commun · 2019

basic_science · Level V

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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.

Medical subject headings