奈米工程與微系統研究所　演講公告

Institute of NanoEngineering and MicroSystems Seminar

2014年12月31日，下午 14:00 – 15:30，台達館 301室

14:00-15:30 pm, December 31, 2014   Room 301, Delta Building

Dr. Lai, Charles Pin-Kuang, Instructor

Harvard Medical School

TITLE:

Visualization and Tracking: Extracellular Vesicles Delivery and RNA Translation Using Multiplexed Reporters

ABSTRACT

Accurate spatiotemporal assessment of extracellular vesicle (EV) delivery and cargo RNA translation requires specific and robust live-cell imaging technologies. Here we engineered optical reporters to label multiple EV populations for visualization and tracking of tumor 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 their NH2-termini with a palmitoylation signal (PalmGFP, PalmtdTomato) for EV membrane labeling. To monitor EV-RNA cargo, transcripts encoding PalmtdTomato were tagged with MS2 RNA binding sequences and detected by coexpression of bacteriophage MS2 coat protein fused with EGFP. By multiplexing fluorescent and bioluminescent EV membrane reporters, we revealed the rapid dynamics of both EV uptake and translation of EV-delivered cargo mRNAs in target cancer cells that occurred within one-hour post-horizontal transfer between cells. We further investigated biodistribution of EVs in vivo and demonstrated accurate spatiotemporal resolution which is previously unachievable. These studies confirm that EV-mediated communication is dynamic and multidirectional between cells with delivery of functional mRNA.

Contact: Chihchen Chen, #62403, chihchen@mx.nthu.edu.tw