1. Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs.
- Author
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Kundert, K, Kundert, K, Lucas, JE, Watters, KE, Fellmann, C, Ng, AH, Heineike, BM, Fitzsimmons, CM, Oakes, BL, Qu, J, Prasad, N, Rosenberg, OS, Savage, DF, El-Samad, H, Doudna, JA, Kortemme, T, Kundert, K, Kundert, K, Lucas, JE, Watters, KE, Fellmann, C, Ng, AH, Heineike, BM, Fitzsimmons, CM, Oakes, BL, Qu, J, Prasad, N, Rosenberg, OS, Savage, DF, El-Samad, H, Doudna, JA, and Kortemme, T
- 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.
- Published
- 2019