Functional genome-wide short hairpin RNA library screening identifies key molecules for extracellular vesicle secretion from microglia.

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⁺ cells with ATP induces rapid secretion of EVs and a reduction in cellular tdTomato intensity, reflecting EV secretion. We generate a GFP⁺ tdTomato⁺ 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

SUBMITTER: Ruan Z 

PROVIDER: S-EPMC9133589 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature