Extracellular vesicles (EVs) are a heterogeneous group of small, lipid-based nanoparticles that play a key role as mediators of many physiological and pathophysiological processes, mainly by the delivery of RNAs. As opposed to the widely known artificial liposomes, EVs are coated with surface proteins that provide specific tissue targeting. This interesting feature, alongside their biocompatibility, bioavailability, and ability to cross the blood-brain barrier, has boosted EVs as a potential RNA drug carrier and lead to multiple therapeutic approaches.
Unfortunately, substantial obstacles block their implementation as therapeutic agents. The ability to produce high amounts of designer EVs and a standard and specific purification methodology are still lacking.
Here we provide a solution for these two issues: we present a biological system to produce designer exosomes based on a cell line, that could be easily scalated.
Firstly, we have included a tag system to improve and simplify the collection and purification techniques, making it more feasible for laboratory groups.
At the same time, we have implemented a molecular device based on L7Ae:C/D-box interaction that permits the direct loading of shRNA labelled molecules into exosomes inside the cell, avoiding the cumbersome process of artificial loading.
Lastly, we pretend to validate our system by producing exosomes loaded with shRNA against myc to use them as an innovative therapy against Burkitt's lymphoma. With that system, that can be adapted to generate different therapeutic exosomes ad libitum, we expect to unleash the potential of engineered exosomes in clinical practice in the future.