- 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
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.