Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors
Journal article, 2025

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.

Author

Xiuming Liang

Karolinska Institutet

Shandong University

Karolinska University Hospital

Dhanu Gupta

University of Oxford

Karolinska Institutet

Junhua Xie

Ghent university

Flanders Interuniversity Institute for Biotechnology

Elien Van Wonterghem

Flanders Interuniversity Institute for Biotechnology

Ghent university

Lien Van Hoecke

Ghent university

Flanders Interuniversity Institute for Biotechnology

Justin Hean

Evox Therapeutics Limited

Zheyu Niu

Karolinska Institutet

Shandong First Medical University

Marziyeh Ghaeidamini

Chalmers, Life Sciences, Chemical Biology

Oscar P.B. Wiklander

Karolinska Institutet

Karolinska University Hospital

Wenyi Zheng

Karolinska Institutet

Karolinska University Hospital

Rim Jawad Wiklander

Karolinska Institutet

Rui He

Karolinska Institutet

Doste R. Mamand

Karolinska Institutet

Karolinska University Hospital

Jeremy P. Bost

Karolinska Institutet

Guannan Zhou

Fudan University

Karolinska Institutet

Houze Zhou

Karolinska Institutet

Karolinska University Hospital

Samantha Roudi

Karolinska Institutet

Karolinska University Hospital

H. Yesid Estupiñán

Karolinska University Hospital

Karolinska Institutet

Industrial University of Santander

Julia Rädler

Karolinska University Hospital

Karolinska Institutet

Antje M. Zickler

Karolinska Institutet

Karolinska University Hospital

André Görgens

Karolinska Institutet

Universitats Klinikum Essen und Medizinische Fakultat

Karolinska University Hospital

Vicky W.Q. Hou

Karolinska University Hospital

Karolinska Institutet

Radka Slovak

Evox Therapeutics Limited

Daniel W. Hagey

Karolinska Institutet

Karolinska University Hospital

Olivier G. de Jong

Utrecht University

Aileen Geobee Uy

Uppsala University

Karolinska Institutet

Yuanyuan Zong

Shandong First Medical University

Imre Mäger

University of Tartu

Carla Martin Perez

University of Oxford

Thomas C. Roberts

University of Oxford

Dave Carter

Evox Therapeutics Limited

Pieter Vader

Utrecht University

Elin Esbjörner Winters

Chalmers, Life Sciences, Chemical Biology

Antonin de Fougerolles

Evox Therapeutics Limited

Matthew J.A. Wood

University of Oxford

Roosmarijn E. Vandenbroucke

Flanders Interuniversity Institute for Biotechnology

Ghent university

Joel Z. Nordin

Karolinska Institutet

Karolinska University Hospital

Samir El-Andaloussi

Karolinska University Hospital

Karolinska Institutet

Nature Communications

2041-1723 (ISSN) 20411723 (eISSN)

Vol. 16 1 4028

Subject Categories (SSIF 2025)

Molecular Biology

Cell Biology

DOI

10.1038/s41467-025-59377-y

PubMed

40301355

More information

Latest update

5/28/2025