PRJNA826851
Genome-wide short-hairpin RNA library screening of murine microglia cell line
Activated microglia release extracellular vesicles (EVs) as modulators of brain homeostasis and innate immunity. However, the molecules critical for regulating the 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 a rapid secretion of EVs and 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.