biostudies-literatureUnknown16(1)Liang XIntracellular 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.Nature communications4028https://www.ebi.ac.uk/biostudies/studies/S-EPMC12041237biostudies-literatureEngineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors.PMC12041237Mamand DRde Fougerolles AWiklander RJSlovak RRoberts TCLiang XGhaeidamini MGorgens ANiu ZBost JHagey DWZickler AMRadler JVan Hoecke LGupta DCarter DHou VWQXie JVader PNordin JZZhou GPerez CMZhou HVan Wonterghem EEstupinan HYZong YWood MJAUy AGMager IZheng WHean JHe Rde Jong OGWiklander OPBEsbjorner EKRoudi SVandenbroucke REEl Andaloussi SfalseEngineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors.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.2025-01-01T00:00:00Z2025 Apr2025-06-25T03:06:21.719Z2025-06-25T03:06:21.719ZS-EPMC120412374030135510.1038/s41467-025-59377-y