Molecular Basis and Regulatory Mechanisms of Exosome Secretion

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ABSTRACT Exosomes are small extracellular vesicles secreted by cells. Exosomes carry bioactive molecules such as signaling proteins and mRNAs, and potently affect the pathophysiology of their recipient cells. The project funded by my R35 grant “Molecular Basis and Regulatory Mechanisms of Exosome Secretion” (R35 GM141832) aims to understanding the molecular mechanisms that control the biogenesis and secretion of exosomes, and the regulation of exosome biogenesis by oncogenic signaling. Due to the small size (30-120 nm in diameter) of exosomes, a nanoparticle tracking analysis (NTA) equipment is essential for the proposed study. The NTA equipment carries out functions ranging from quantification of exosome number, measuring exosome diameters, to fluorescence-based cargo labeling and exosome sub-population identification. Without these basic exosome characterizations, most of our proposed experiments cannot be carried out.In this proposal, I apply for Administrative Supplements for Equipment fund (NOT-GM-22-017) for the purchase of a ZetaView® x30 dual laser multiple parameter NTA device to replace our broken equipment, NanoSight N300. We plan to purchase a ZetaView®x30 from Particle Metrix, Inc. rather than the NanoSight NS300 from Malvin, Inc. because ZetaView®x30 is a leading NTA equipment in the market with features meeting our research needs, whereas the old NanoSight NS300 system has a number of deficiencies revealed over these years. In fact, ZetaView system is the only NTA device that allow us to characterize exosomes with precision, and is indispensable for the accomplishment of the goals proposed in the parent R35 grant.