Abstract
Extracellular vesicles (EVs) play a key role in cancer development and cellular homeostasis by transferring the biological cargo to recipient cells. Here, we describe steps for screening EV secretion-related genes by combining a microRNA (miRNA) library and ExoScreen, a highly sensitive EV detection technique. We also detail procedures for screening the direct target genes regulated by miRNAs. This protocol provides a useful tool for understanding complex intracellular communications involved in EV secretion. For complete details on the use and execution of this protocol, please refer to Yamamoto et al.1.