Isolation of extracellular vesicles and characterization of their role as shuttle of biologically active small RNAs in intercellular communication

Cell-cell communication is crucial for the coordination of various biological processes in multicellular organisms and is mediated by multiple factors, such as hormones, peptides, and small non-coding RNAs. More recently, another mode of intercellular communication has emerged through the encapsulation of biologically active components in extracellular vesicles (EVs). EVs have attracted interest not only as mediators of communication between neighboring cells, but also for their role in cross-kingdom communication between interacting organisms. In this work, we present an optimized protocol for the isolation of EVs from Nicotiana benthamiana leaves and the preliminary characterization of the small RNA composition of EVs.