RT Journal Article
SR Electronic
T1 A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery
JF Life Science Alliance
JO Life Sci. Alliance
FD Life Science Alliance LLC
SP e202201715
DO 10.26508/lsa.202201715
VO 6
IS 4
A1 Patrick Essletzbichler
A1 Vitaly Sedlyarov
A1 Fabian Frommelt
A1 Didier Soulat
A1 Leonhard X Heinz
A1 Adrijana Stefanovic
A1 Benedikt Neumayer
A1 Giulio Superti-Furga
YR 2023
UL https://www.life-science-alliance.org/content/6/4/e202201715.abstract
AB Phagocytosis, the process by which cells engulf large particles, plays a vital role in driving tissue clearance and host defense. Its dysregulation is connected to autoimmunity, toxic accumulation of proteins, and increased risks for infections. Despite its importance, we lack full understanding of all molecular components involved in the process. To create a functional map in human cells, we performed a genome-wide CRISPRko FACS screen that identified 716 genes. Mapping those hits to a comprehensive protein–protein interaction network annotated for functional cellular processes allowed retrieval of protein complexes identified multiple times and detection of missing phagocytosis regulators. In addition to known components, such as the Arp2/3 complex, the vacuolar-ATPase-Rag machinery, and the Wave-2 complex, we identified and validated new phagocytosis-relevant functions, including the oligosaccharyltransferase complex (MAGT1/SLC58A1, DDOST, STT3B, and RPN2) and the hypusine pathway (eIF5A, DHPS, and DOHH). Overall, our phagocytosis network comprises elements of cargo uptake, shuffling, and biotransformation through the cell, providing a resource for the identification of potential novel drivers for diseases of the endo-lysosomal system. Our approach of integrating protein–protein interaction offers a broadly applicable way to functionally interpret genome-wide screens.