{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Ruan Z"],"funding":["NIA NIH HHS"],"pagination":["110791"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9133589"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["39(6)"],"pubmed_abstract":["Activated microglia release extracellular vesicles (EVs) as modulators of brain homeostasis and innate immunity. However, the molecules critical for regulating EV production from microglia are poorly understood. Here we establish a murine microglial cell model to monitor EV secretion by measuring the fluorescence signal of tdTomato, which is linked to tetraspanin CD63. Stimulation of tdTomato<sup>+</sup> cells with ATP induces rapid secretion of EVs and a reduction in cellular tdTomato intensity, reflecting EV secretion. We generate a GFP<sup>+</sup> tdTomato<sup>+</sup> cell library expressing TurboGFP and barcoded short hairpin RNAs for genome-wide screening using next-generation sequencing. We identify Mcfd2, Sepp1, and Sdc1 as critical regulators of ATP-induced EV secretion from murine"],"journal":["Cell reports"],"pubmed_title":["Functional genome-wide short hairpin RNA library screening identifies key molecules for extracellular vesicle secretion from microglia."],"pmcid":["PMC9133589"],"funding_grant_id":["P30 AG013846","R01 AG067763","R01 AG054672","R01 AG054199","R01 AG072719","RF1 AG054199","R01 AG066429"],"pubmed_authors":["Ruan Z","Takamatsu-Yukawa K","Ikezu S","Ikezu T","Labadorf AT","Wang Y","Ericsson M","Ushman ML"],"additional_accession":[]},"is_claimable":false,"name":"Functional genome-wide short hairpin RNA library screening identifies key molecules for extracellular vesicle secretion from microglia.","description":"Activated microglia release extracellular vesicles (EVs) as modulators of brain homeostasis and innate immunity. However, the molecules critical for regulating EV production from microglia are poorly understood. Here we establish a murine microglial cell model to monitor EV secretion by measuring the fluorescence signal of tdTomato, which is linked to tetraspanin CD63. Stimulation of tdTomato<sup>+</sup> cells with ATP induces rapid secretion of EVs and a reduction in cellular tdTomato intensity, reflecting EV secretion. We generate a GFP<sup>+</sup> tdTomato<sup>+</sup> cell library expressing TurboGFP and barcoded short hairpin RNAs for genome-wide screening using next-generation sequencing. We identify Mcfd2, Sepp1, and Sdc1 as critical regulators of ATP-induced EV secretion from murine","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 May","modification":"2026-05-09T18:55:30.269Z","creation":"2025-05-18T12:06:19.777Z"},"accession":"S-EPMC9133589","cross_references":{"pubmed":["35545052"],"doi":["10.1016/j.celrep.2022.110791"]}}