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Nat Commun. 2019 May 9;10(1):2127. doi: 10.1038/s41467-019-09985-2.

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

Author information

1
Graduate Group in Biophysics, University of California San Francisco, San Francisco, CA, 94158, USA. kale@thekunderts.net.
2
UC Berkeley - UCSF Graduate Program in Bioengineering, University of California San Francisco, San Francisco, CA, 94158, USA.
3
Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, 94704, USA.
4
Bioinformatics Graduate Program, University of California San Francisco, San Francisco, CA, 94158, USA.
5
Chemistry and Chemical Biology Graduate Program, University of California San Francisco, San Francisco, CA, 94158, USA.
6
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
7
Department of Medicine, University of California San Francisco, San Francisco, CA, 94158, USA.
8
Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA.
9
Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA, 94158, USA.
10
Howard Hughes Medical Institute, University of California Berkeley, Berkeley, CA, 94704, USA.
11
Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA. tanjakortemme@gmail.com.
12
Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA, 94158, USA. tanjakortemme@gmail.com.

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.

PMID:
31073154
PMCID:
PMC6509140
DOI:
10.1038/s41467-019-09985-2
[Indexed for MEDLINE]
Free PMC Article

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