Today (Wednesday, June 22), in the last of four Featured Abstracts presented by graduate students during the ISEV 2020 Virtual Annual Meeting (July 20-22) (https://www.eventscribe.com/2020/ISEV/), Bhagyashree Joshi (photo), of the Department of Biomedical Engineering, University of Groningen, University Medical Center Groningen, The Netherlands, presented her group’s abstract (FA03) “Genetically Encoded Probes Provide Insight into Extracellular Vesicle Cargo Release in Cells.” Ms. Joshi is a PhD candidate in the laboratory of Inge Zuhorn, PhD, Associate Professor, Department of Biomedical Engineering, University of Groningen, University Medical Center Groningen University Medical Center Groningen. In her introduction, Ms. Joshi noted that extracellular vesicles (EVs) are known to modulate tissue development, regeneration, and disease through the transfer of proteins, nucleic acids, and lipids between cells. Currently, however, the mechanism of cytosolic delivery of EV cargo is largely unknown, she said. It has been speculated that EVs undergo back fusion at multi-vesicular bodies (MVBs) in recipient cells to release their functional cargo. However, Ms. Joshi said, evidence for this is lacking. She remarked that tracing the cellular uptake of EVs with high resolution, as well as acquiring direct evidence for the release of EV cargo, is challenging, chiefly because of technical limitations. To address this problem, Ms. Joshi and colleagues developed an analytical methodology that combined state-of-the-art molecular tools and correlative light and electron microscopy (CLEM) to identify the intracellular site for EV cargo release. Green fluorescent protein (GFP) was loaded inside EVs through the expression of GFP-CD63, a fusion of GFP to the cytosolic tail of CD63, in EV producer cells.
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