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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 importance, we lack full understanding of...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Life Science Alliance LLC
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9892931/ https://www.ncbi.nlm.nih.gov/pubmed/36725334 http://dx.doi.org/10.26508/lsa.202201715 |
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author | Essletzbichler, Patrick Sedlyarov, Vitaly Frommelt, Fabian Soulat, Didier Heinz, Leonhard X Stefanovic, Adrijana Neumayer, Benedikt Superti-Furga, Giulio |
author_facet | Essletzbichler, Patrick Sedlyarov, Vitaly Frommelt, Fabian Soulat, Didier Heinz, Leonhard X Stefanovic, Adrijana Neumayer, Benedikt Superti-Furga, Giulio |
author_sort | Essletzbichler, Patrick |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9892931 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Life Science Alliance LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-98929312023-02-03 A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery Essletzbichler, Patrick Sedlyarov, Vitaly Frommelt, Fabian Soulat, Didier Heinz, Leonhard X Stefanovic, Adrijana Neumayer, Benedikt Superti-Furga, Giulio Life Sci Alliance Research Articles 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. Life Science Alliance LLC 2023-02-01 /pmc/articles/PMC9892931/ /pubmed/36725334 http://dx.doi.org/10.26508/lsa.202201715 Text en © 2023 Essletzbichler et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Articles Essletzbichler, Patrick Sedlyarov, Vitaly Frommelt, Fabian Soulat, Didier Heinz, Leonhard X Stefanovic, Adrijana Neumayer, Benedikt Superti-Furga, Giulio A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery |
title | A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery |
title_full | A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery |
title_fullStr | A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery |
title_full_unstemmed | A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery |
title_short | A genome-wide CRISPR functional survey of the human phagocytosis molecular machinery |
title_sort | genome-wide crispr functional survey of the human phagocytosis molecular machinery |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9892931/ https://www.ncbi.nlm.nih.gov/pubmed/36725334 http://dx.doi.org/10.26508/lsa.202201715 |
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