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Scalable, FACS-Free Genome-Wide Phenotypic Screening

By Barbara Mair, Peter M. Aldridge, Randy S. Atwal, Sanna N. Masud, Meng Zhang, David Philpott, Amy H.Y. Tong, Edward H. Sargent, Stephane Angers, Jason Moffat, Shana O. Kelley

Posted 19 Apr 2019
bioRxiv DOI: 10.1101/612887

Genome-scale functional genetic screens can identify key regulators of a phenotype of interest, such as determinants of protein expression or modification. Here, we present a rapid, high-throughput approach to phenotypic CRISPR-Cas9 screening. To study factors that modulate the display of CD47 on the cell surface, we processed an entire genome-wide screen containing more than 108 cells in under one hour and maintained high levels of cell viability using a highly scalable cell sorting technology. We robustly identified modulators of CD47 function including QPCTL, an enzyme required for formation of the pyroglutamyl modification at the N-terminus of this protein.

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