Fluorogenic RNA-based biomaterials for imaging and tracking the cargo of extracellular vesicles

Area of Science:

• Molecular Biology
• Cell Biology
• Biotechnology

Background:

• Extracellular vesicles (EVs) are crucial for cell communication and are promising as drug carriers.
• EVs contain non-coding RNAs that regulate gene expression and epigenetic processes.
• Understanding EV RNA trafficking is key to optimizing their therapeutic applications.

Purpose of the Study:

• To develop a tool for visualizing and tracking RNA within EVs.
• To investigate the role of EVs in RNA-based cellular communication.
• To enhance the therapeutic potential of EVs.

Main Methods:

• Development of EXO-Probe, a genetically encodable RNA construct.
• EXO-Probe fuses an EV-loading RNA sequence (EXO-Code) with a fluorogenic RNA Mango aptamer.
• Utilized the system for RNA visualization, tracking, colocalization studies, and non-destructive EV quantification.

Main Results:

• Successfully visualized and tracked EV RNAs using EXO-Probe.
• Demonstrated colocalization of EXO-Probe with multivesicular body markers.
• Enabled non-destructive quantification of EVs and imaging of RNA within them.

Conclusions:

• EXO-Probe is a versatile tool for studying EV RNA trafficking and cellular sorting.
• Provides insights into EV-mediated RNA communication.
• Lays the groundwork for improving the therapeutic efficacy of EVs.