Scalable, FACS-Free Genome-Wide Phenotypic Screening

Mair, B.; Aldridge, P. M.; Atwal, R. S.; Masud, S. N.; Zhang, M.; Philpott, D.; Tong, A. H. Y.; Sargent, E. H.; Angers, S.; Moffat, J.; Kelley, S. O.

doi:10.1101/612887

bioRxiv · 10.1101/612887

Scalable, FACS-Free Genome-Wide Phenotypic Screening

Mair, B.·Aldridge, P. M.·Atwal, R. S.·Masud, S. N.·Zhang, M.·Philpott, D.·Tong, A. H. Y.·Sargent, E. H.·Angers, S.·Moffat, J.·Kelley, S. O.

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Abstract

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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Mair, B., Aldridge, P. M., Atwal, R. S., Masud, S. N., Zhang, M., Philpott, D., Tong, A. H. Y., Sargent, E. H., Angers, S., Moffat, J., Kelley, S. O.. 2019-04-19. Scalable, FACS-Free Genome-Wide Phenotypic Screening. https://doi.org/10.1101/612887

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