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

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.

SUBMITTER: Kundert K

PROVIDER: S-EPMC6509140 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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

Kundert Kale K Lucas James E JE Watters Kyle E KE Fellmann Christof C Ng Andrew H AH Heineike Benjamin M BM Fitzsimmons Christina M CM Oakes Benjamin L BL Qu Jiuxin J Prasad Neha N Rosenberg Oren S OS Savage David F DF El-Samad Hana H Doudna Jennifer A JA Kortemme Tanja T

Nature communications 20190509 1

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 resp ...[more]

PMID: 31073154

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