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

Kundert, Kale; Lucas, James E.; Watters, Kyle E.; Fellmann, Christof; Ng, Andrew H.; Heineike, Benjamin M.; Fitzsimmons, Christina M.; Oakes, Benjamin L.; Qu, Jiuxin; Prasad, Neha; Rosenberg, Oren S.; Savage, David F.; El-Samad, Hana; Doudna, Jennifer A.; Kortemme, Tanja

doi:10.1038/s41467-019-09985-2

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

Journal Article · Thu May 09 00:00:00 EDT 2019 · Nature Communications

DOI:https://doi.org/10.1038/s41467-019-09985-2· OSTI ID:1603511

^[1]; Lucas, James E. ^[2]; ^[3]; ^[3]; ^[4]; Heineike, Benjamin M. ^[1]; Fitzsimmons, Christina M. ^[5]; Oakes, Benjamin L. ^[3]; Qu, Jiuxin ^[1]; Prasad, Neha ^[1]; ^[6]; Savage, David F. ^[3]; El-Samad, Hana ^[7]; ^[8]; ^[7]

Univ. of California, San Francisco, CA (United States)
Univ. of California, San Francisco, CA (United States). UC Berkeley – UCSF Graduate Program in Bioengineering
Univ. of California, Berkeley, CA (United States)
Univ. of California, San Francisco, CA (United States). UC Berkeley – UCSF Graduate Program in Bioengineering
Univ. of California, San Francisco, CA (United States); National Inst. of Health (NIH), Bethesda, MD (United States). National Cancer Inst.
Univ. of California, San Francisco, CA (United States); Chan Zuckerberg Biohub, San Francisco, CA (United States)
Chan Zuckerberg Biohub, San Francisco, CA (United States); Univ. of California, San Francisco, CA (United States)
Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst.

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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1603511

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 10; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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