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

Kundert, K.; Lucas, J. E.; Watters, K. E.; Fellmann, C.; Ng, A. H.; Heineike, B. M.; Fitzsimmons, C. M.; Oakes, B. L.; Savage, D. F.; El-Samad, H.; Doudna, J. A.; Kortemme, T.

doi:10.1101/323105

bioRxiv · 10.1101/323105

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

Kundert, K.·Lucas, J. E.·Watters, K. E.·Fellmann, C.·Ng, A. H.·Heineike, B. M.·Fitzsimmons, C. M.·Oakes, B. L.·Savage, D. F.·El-Samad, H.·Doudna, J. A.·Kortemme, T.

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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.

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Kundert, K., Lucas, J. E., Watters, K. E., Fellmann, C., Ng, A. H., Heineike, B. M., Fitzsimmons, C. M., Oakes, B. L., Savage, D. F., El-Samad, H., Doudna, J. A., Kortemme, T.. 2018-05-15. Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs. https://doi.org/10.1101/323105

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