Fluorogenic EXO-Probe aptamers for imaging and tracking exosomal RNAs

EE Bonacquisti, SW Ferguson, NE Jasiewicz, J Wang… - bioRxiv, 2021 - biorxiv.org
bioRxiv, 2021biorxiv.org
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 …
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.
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