A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery

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 im-portance, 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 iden-tified 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 in-terpret genome-wide screens.

Automated summary

Phagocytosis is a crucial process for tissue clearance and host defense, but its dysregulation is linked to various diseases. This study used a genome-wide CRISPRko FACS screen to identify 716 genes involved in phagocytosis. By mapping the hits to a protein-protein interaction network, novel phagocytosis regulators were discovered and validated. This comprehensive network provides insights into cargo uptake and transformation, and can aid in the identification of potential drivers for endo-lysosomal diseases.