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
Karolinska University Hospital
Shandong University
Dhanu Gupta
Karolinska Institutet
University of Oxford
Junhua Xie
Flanders Interuniversity Institute for Biotechnology
Ghent university
Elien Van Wonterghem
Ghent university
Flanders Interuniversity Institute for Biotechnology
Lien Van Hoecke
Flanders Interuniversity Institute for Biotechnology
Ghent university
Justin Hean
Evox Therapeutics Limited
Zheyu Niu
Shandong First Medical University
Karolinska Institutet
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
Karolinska Institutet
Obstetrics & Gynecology Hospital of Fudan University
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 Institutet
Karolinska University Hospital
Antje M. Zickler
Karolinska University Hospital
Karolinska Institutet
André Görgens
Karolinska University Hospital
Karolinska Institutet
Universitats Klinikum Essen und Medizinische Fakultat
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
Utrechts Instituut voor Farmaceutische Wetenschappen
Aileen Geobee Uy
Karolinska Institutet
Uppsala University
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
University Medical Center Utrecht
Elin Esbjörner Winters
Chalmers, Life Sciences, Chemical Biology
Antonin de Fougerolles
Evox Therapeutics Limited
Matthew J.A. Wood
University of Oxford
Roosmarijn E. Vandenbroucke
Ghent university
Flanders Interuniversity Institute for Biotechnology
Joel Z. Nordin
Karolinska Institutet
Karolinska University Hospital
Samir El-Andaloussi
Karolinska Institutet
Karolinska University Hospital
Nature Communications
2041-1723 (ISSN) 20411723 (eISSN)
Vol. 16 1 4028Subject Categories (SSIF 2025)
Molecular Biology
Cell Biology
DOI
10.1038/s41467-025-59377-y