RT Journal Article
SR Electronic
T1 Decoding directional genetic dependencies through orthogonal CRISPR/Cas screens
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 120170
DO 10.1101/120170
A1 Michael Boettcher
A1 Ruilin Tian
A1 James Blau
A1 Evan Markegard
A1 David Wu
A1 Anne Biton
A1 Noah Zaitlen
A1 Frank McCormick
A1 Martin Kampmann
A1 Michael T. McManus
YR 2017
UL http://biorxiv.org/content/early/2017/03/25/120170.abstract
AB Genetic interaction studies are a powerful approach to identify functional interactions between genes. This approach can reveal networks of regulatory hubs and connect uncharacterised genes to well-studied pathways. However, this approach has previously been limited to simple gene inactivation studies. Here, we present an orthogonal CRISPR/Cas-mediated genetic interaction approach that allows the systematic activation of one gene while simultaneously knocking out a second gene in the same cell. We have developed this concept into a quantitative and scalable combinatorial screening platform that allows the parallel interrogation of hundreds of thousands of genetic interactions. We demonstrate that the established platform works robustly to uncover genetic interactions in human cancer cells and to interpret the direction of the flow of genetic information.