Visualization and tracking of tumour extracellular vesicle delivery and RNA translation using multiplexed reporters

Accurate spatiotemporal assessment of extracellular vesicle (EV) delivery and cargo RNA translation requires specific and robust live-cell imaging technologies. Here we engineer optical reporters to label multiple EV populations for visualization and tracking of tumour EV release, uptake and exchange between cell populations both in culture and in vivo. Enhanced green fluorescence protein (EGFP) and tandem dimer Tomato (tdTomato) were fused at NH2-termini with a palmitoylation signal (PalmGFP, PalmtdTomato) for EV membrane labelling. To monitor EV-RNA cargo, transcripts encoding PalmtdTomato were tagged with MS2 RNA binding sequences and detected by co-expression of bacteriophage MS2 coat protein fused with…

• VA
  Voices Against Brain Cancer
• NI
  National Institutes of Health

  Awards: U19CA179563, 1S10RR023440-01A1, CA069246, P30NS045776, CA179563
• CI
  Canadian Institutes of Health Research
• NO
  NIH Office of the Director

  Awards: CA179563, U19CA179563
• NI
  National Institute of Neurological Disorders and Stroke

  Award: P30NS045776
• OO
  Office of Strategic Coordination
• CF
  Common Fund