Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors
Publication date
2025-04-29
Authors
Liang, Xiuming
Gupta, Dhanu
Xie, Junhua
Van Wonterghem, Elien
Van Hoecke, Lien
Hean, Justin
Niu, Zheyu
Ghaeidamini, Marziyeh
Wiklander, Oscar P B
Zheng, Wenyi
Editors
Advisors
Supervisors
Document Type
Article
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cc_by
Abstract
Intracellular delivery of protein and RNA therapeutics represents a major challenge. Here, we develop highly potent engineered extracellular vesicles (EVs) by incorporating bio-inspired attributes required for effective delivery. These comprise an engineered mini-intein protein with self-cleavage activity for active cargo loading and release, and fusogenic VSV-G protein for endosomal escape. Combining these components allows high efficiency recombination and genome editing in vitro following EV-mediated delivery of Cre recombinase and Cas9/sgRNA RNP cargoes, respectively. In vivo, infusion of a single dose Cre loaded EVs into the lateral ventricle in brain of Cre-LoxP R26-LSL-tdTomato reporter mice results in greater than 40% and 30% recombined cells in hippocampus and cortex respectively. In addition, we demonstrate therapeutic potential of this platform by showing inhibition of LPS-induced systemic inflammation via delivery of a super-repressor of NF-ĸB activity. Our data establish these engineered EVs as a platform for effective delivery of multimodal therapeutic cargoes, including for efficient genome editing.
Keywords
Animals, CRISPR-Associated Protein 9/metabolism, CRISPR-Cas Systems, Drug Delivery Systems/methods, Extracellular Vesicles/metabolism, Gene Editing/methods, HEK293 Cells, Humans, Inflammation/chemically induced, Integrases/genetics, Mice, NF-kappa B/metabolism, Journal Article
Citation
Liang, X, Gupta, D, Xie, J, Van Wonterghem, E, Van Hoecke, L, Hean, J, Niu, Z, Ghaeidamini, M, Wiklander, O P B, Zheng, W, Wiklander, R J, He, R, Mamand, D R, Bost, J, Zhou, G, Zhou, H, Roudi, S, Estupiñán, H Y, Rädler, J, Zickler, A M, Görgens, A, Hou, V W Q, Slovak, R, Hagey, D W, de Jong, O G, Uy, A G, Zong, Y, Mäger, I, Perez, C M, Roberts, T C, Carter, D, Vader, P, Esbjörner, E K, de Fougerolles, A, Wood, M J A, Vandenbroucke, R E, Nordin, J Z & El Andaloussi, S 2025, 'Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors', Nature Communications, vol. 16, no. 1, 4028. https://doi.org/10.1038/s41467-025-59377-y