Affiliation:
1. Karolinska Institutet
2. Ghent University
3. Evox Therapeutics Limited
4. Shandong First Medical University
5. Scibase Company
6. Utrecht University
7. Uppsala University
8. University of Tartu
9. University of Oxford
10. University Medical Center Utrecht
Abstract
Abstract
Intracellular delivery of protein and RNA therapeutics represents a major challenge. Here, we developed highly potent engineered extracellular vesicles (EVs) by incorporating essential bio-inspired attributes required for effective delivery. These comprise engineered mini-intein proteins with self-cleavage activity for active cargo loading and release, and fusogenic VSV-G protein to activate productive endosomal escape. Combining these components allowed high efficiency recombination and genome editing in vitrofollowing EV-mediated delivery of Cre recombinase and Cas9/sgRNA RNP cargoes, respectively. In vivo, single dose EV-mediated Cre delivery to the brains of Cre-LoxP R26-LSL-tdTomato reporter mice resulted 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 novel platform for effective delivery of multimodal therapeutic cargoes, including for efficient genome editing.
Publisher
Research Square Platform LLC