Scalable, FACS-Free Genome-Wide Phenotypic Screening
Peter M. Aldridge,
Randy S. Atwal,
Sanna N. Masud,
Amy H.Y. Tong,
Edward H. Sargent,
Shana O. Kelley
Posted 19 Apr 2019
bioRxiv DOI: 10.1101/612887
Posted 19 Apr 2019
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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