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

Charles P. Lai (), Edward Y. Kim, Christian E. Badr, Ralph Weissleder, Thorsten R. Mempel, Bakhos A. Tannous and Xandra O. Breakefield ()
Additional contact information
Charles P. Lai: Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School
Edward Y. Kim: Massachusetts General Hospital, and Program in Immunology, Harvard Medical School
Christian E. Badr: Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School
Ralph Weissleder: Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School
Thorsten R. Mempel: Massachusetts General Hospital, and Program in Immunology, Harvard Medical School
Bakhos A. Tannous: Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School
Xandra O. Breakefield: Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School

Nature Communications, 2015, vol. 6, issue 1, 1-12

Abstract: Abstract 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 EGFP. By multiplexing fluorescent and bioluminescent EV membrane reporters, we reveal the rapid dynamics of both EV uptake and translation of EV-delivered cargo mRNAs in cancer cells that occurred within 1-hour post-horizontal transfer between cells. These studies confirm that EV-mediated communication is dynamic and multidirectional between cells with delivery of functional mRNA.

Date: 2015
References: Add references at CitEc
Citations: View citations in EconPapers (2)

Downloads: (external link)
https://www.nature.com/articles/ncomms8029 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8029

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/ncomms8029

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().