Extracellular Vesicles Enable CircRNA Delivery via in situ Biogenesis and Sorting

Abstract

Extracellular vesicles (EVs) are promising vehicles for nucleic acid delivery, yet efficient delivery of circular RNA (circRNA) remains challenging due to inefficient loading and limited intracellular expression. Here, we establish an EV-based platform that enables efficient circRNA delivery via in situ biogenesis and sorting. By optimizing intracellular circularization and translation through vector design, we markedly enhance circRNA expression. By combining this with Snu13-mediated EV sorting and enhanced vesicle biogenesis, we achieve efficient packaging of circRNA without compromising vesicle integrity. This integrated strategy enables robust and sustained protein expression following EV-based circRNA delivery. By leveraging this platform, we demonstrate a dendritic cell-targeting circRNA vaccine that elicits strong antigen-specific CD8+ T cell responses and antitumor efficacy. We further show that systemic delivery of BNP-encoding circRNA attenuates doxorubicin-induced myocardial fibrosis. Together, this work establishes a generalizable platform for circRNA therapeutics by overcoming key barriers in circRNA expression and EV-mediated delivery. TeaserEngineered EVs enable efficient circRNA delivery for sustained protein expression and therapy.