RT Journal Article
SR Electronic
T1 Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 323105
DO 10.1101/323105
A1 Kale Kundert
A1 James E. Lucas
A1 Kyle E. Watters
A1 Christof Fellmann
A1 Andrew H. Ng
A1 Benjamin M. Heineike
A1 Christina M. Fitzsimmons
A1 Benjamin L. Oakes
A1 David F. Savage
A1 Hana El-Samad
A1 Jennifer A. Doudna
A1 Tanja Kortemme
YR 2018
UL http://biorxiv.org/content/early/2018/05/15/323105.abstract
AB 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.