Fluorogenic EXO-Probe Aptamers for Imaging and Tracking Exosomal RNAs
bioRxiv(2021)
Abstract
Small extracellular vesicles (sEVs), or exosomes, play important roles in physiological and pathological cellular communication. sEVs contain both short and long non-coding RNAs that regulate gene expression and epigenetic processes. Studying the intricacies of sEV function and RNA-based communication requires tools capable of labeling sEV RNA. Here we developed a novel genetically encodable reporter system for tracking sEV RNAs comprising an sEV-loading RNA sequence, termed the EXO-Code, fused to a fluorogenic RNA Mango aptamer for RNA imaging. This fusion construct allowed the visualization and tracking of RNA puncta and colocalization with markers of multivesicular bodies; imaging RNA puncta within sEVs; and quantification of sEVs. This technology represents a useful and versatile tool to interrogate the role of sEVs in cellular communication via RNA trafficking to sEVs, cellular sorting decisions, and sEV RNA cargo transfer to recipient cells.
### Competing Interest Statement
JN, SF, and EB are inventors on the patent applications of the EXO-Code technology evaluated in this paper that have been licensed to Exopharm and have received royalties. These relationships have been disclosed to and are under management by UNC-Chapel Hill.
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Key words
imaging,exo-probe
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