Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens

Michael S. Haney, Christopher J. Bohlen, David W. Morgens, James A. Ousey, Amira A. Barkal, C. Kimberly Tsui, Braeden K. Ego, Roni Levin, Roarke A. Kamber, Hannah Collins, Andrew Tucker, Amy Li, Daan Vorselen, Lorenzo Labitigan, Emily Crane, Evan Boyle, Lihua Jiang, Joanne Chan, Esther Rincón, William J. GreenleafBilly Li, Michael P. Snyder, Irving L. Weissman, Julie A. Theriot, Sean Collins, Ben A. Barres, Michael C. Bassik

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

Phagocytosis is required for a broad range of physiological functions, from pathogen defense to tissue homeostasis, but the mechanisms required for phagocytosis of diverse substrates remain incompletely understood. Here, we developed a rapid magnet-based phenotypic screening strategy, and performed eight genome-wide CRISPR screens in human cells to identify genes regulating phagocytosis of distinct substrates. After validating select hits in focused miniscreens, orthogonal assays and primary human macrophages, we show that (1) the previously uncharacterized gene NHLRC2 is a central player in phagocytosis, regulating RhoA-Rac1 signaling cascades that control actin polymerization and filopodia formation, (2) very-long-chain fatty acids are essential for efficient phagocytosis of certain substrates and (3) the previously uncharacterized Alzheimer’s disease–associated gene TM2D3 can preferentially influence uptake of amyloid-β aggregates. These findings illuminate new regulators and core principles of phagocytosis, and more generally establish an efficient method for unbiased identification of cellular uptake mechanisms across diverse physiological and pathological contexts.

Original languageEnglish (US)
JournalNature Genetics
DOIs
StateAccepted/In press - Jan 1 2018

ASJC Scopus subject areas

  • Genetics

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    Haney, M. S., Bohlen, C. J., Morgens, D. W., Ousey, J. A., Barkal, A. A., Tsui, C. K., Ego, B. K., Levin, R., Kamber, R. A., Collins, H., Tucker, A., Li, A., Vorselen, D., Labitigan, L., Crane, E., Boyle, E., Jiang, L., Chan, J., Rincón, E., ... Bassik, M. C. (Accepted/In press). Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens. Nature Genetics. https://doi.org/10.1038/s41588-018-0254-1