Programmable RNA Loading of Extracellular Vesicles with Toehold-Release Purification

Mette Galsgaard Malle, Ping Song, Philipp M.G. Löffler, Nazmie Kalisi, Yan Yan, Julián Valero, Stefan Vogel*, Jørgen Kjems*

*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

1 Citationer (Scopus)

Abstract

Synthetic nanoparticles as lipid nanoparticles (LNPs) are widely used as drug delivery vesicles. However, they hold several drawbacks, including low biocompatibility and unfavorable immune responses. Naturally occurring extracellular vesicles (EVs) hold the potential as native, safe, and multifunctional nanovesicle carriers. However, loading of EVs with large biomolecules remains a challenge. Here, we present a controlled loading methodology using DNA-mediated and programmed fusion between EVs and messenger RNA (mRNA)-loaded liposomes. The fusion efficiency is characterized at the single-particle level by real-time microscopy through EV surface immobilization via lipidated biotin-DNA handles. Subsequently, fused EV−liposome particles (EVLs) can be collected by employing a DNA strand-replacement reaction. Transferring the fusion reaction to magnetic beads enables us to scale up the production of EVLs one million times. Finally, we demonstrated encapsulation of mCherry mRNA, transfection, and improved translation using the EVLs compared to liposomes or LNPs in HEK293-H cells. We envision this as an important tool for the EV-mediated delivery of RNA therapeutics.

OriginalsprogEngelsk
TidsskriftJournal of the American Chemical Society
Vol/bind146
Nummer18
Sider (fra-til)12410-12422
Antal sider13
ISSN0002-7863
DOI
StatusUdgivet - maj 2024

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