Functional genome-wide short hairpin RNA library screening identifies key molecules for extracellular vesicle secretion from microglia
Zhi Ruan, Kayo Takamatsu-Yukawa, Yuzhi Wang, Margaret Ushman, Adam Labadorf, Maria Ericsson, Seiko Ikezu, Tsuneya Ikezu
Abstract
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 microglia. Small interfering RNA (siRNA-based) silencing of each of these genes suppresses lipopolysaccharide- and ATP-induced inflammasome activation, as determined by interleukin-1β release from primary cultured murine microglia. These molecules are critical for microglial EV secretion and are potential therapeutic targets for neuroinflammatory disorders.
Topics & Concepts
MicrogliaSecretionSmall hairpin RNAExtracellular vesicleMicrovesiclesCell biologyExtracellular vesiclesRNAGenomeBiologyExtracellularChemistryComputational biologyGeneInflammationmicroRNABiochemistryImmunologyExtracellular vesicles in diseaseNeuroinflammation and Neurodegeneration MechanismsRNA regulation and disease