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 language | English (US) |
|---|---|
| Journal | Nature Genetics |
| DOIs | |
| State | Accepted/In press - Jan 1 2018 |
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ASJC Scopus subject areas
- Genetics
Cite this
Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens. / Haney, Michael S.; Bohlen, Christopher J.; Morgens, David W.; Ousey, James A.; Barkal, Amira A.; Tsui, C. Kimberly; Ego, Braeden K.; Levin, Roni; Kamber, Roarke A.; Collins, Hannah; Tucker, Andrew; Li, Amy; Vorselen, Daan; Labitigan, Lorenzo; Crane, Emily; Boyle, Evan; Jiang, Lihua; Chan, Joanne; Rincón, Esther; Greenleaf, William J.; Li, Billy; Snyder, Michael P.; Weissman, Irving L.; Theriot, Julie A.; Collins, Sean; Barres, Ben A.; Bassik, Michael C.
In: Nature Genetics, 01.01.2018.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens
AU - Haney, Michael S.
AU - Bohlen, Christopher J.
AU - Morgens, David W.
AU - Ousey, James A.
AU - Barkal, Amira A.
AU - Tsui, C. Kimberly
AU - Ego, Braeden K.
AU - Levin, Roni
AU - Kamber, Roarke A.
AU - Collins, Hannah
AU - Tucker, Andrew
AU - Li, Amy
AU - Vorselen, Daan
AU - Labitigan, Lorenzo
AU - Crane, Emily
AU - Boyle, Evan
AU - Jiang, Lihua
AU - Chan, Joanne
AU - Rincón, Esther
AU - Greenleaf, William J.
AU - Li, Billy
AU - Snyder, Michael P.
AU - Weissman, Irving L.
AU - Theriot, Julie A.
AU - Collins, Sean
AU - Barres, Ben A.
AU - Bassik, Michael C.
PY - 2018/1/1
Y1 - 2018/1/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85055975083&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85055975083&partnerID=8YFLogxK
U2 - 10.1038/s41588-018-0254-1
DO - 10.1038/s41588-018-0254-1
M3 - Article
C2 - 30397336
AN - SCOPUS:85055975083
JO - Nature Genetics
JF - Nature Genetics
SN - 1061-4036
ER -