The project focused on developing next-generation gene delivery vectors. Leveraging the natural biocompatibility and cellular uptake efficiency of EVs, we design EV-based and synthetic nanovesicle systems for therapeutic nucleic acid delivery. We have engineered a membrane-enveloped AAV nanovesicle (EV-AAV) that markedly enhances cellular entry, transgene expression, and resistance to antibody neutralization compared to conventional recombinant AAV (rAAV). In animal models of ischemic myocardial injury, hepatocellular carcinoma, and skin wound regeneration, EV-AAV demonstrated superior therapeutic outcomes. Building on this concept, we have recently developed artificial nanovesicles loaded with AAV (CME-AAV) using microfluidic and extrusion-based techniques, achieving higher production efficiency and improved therapeutic efficacy in a Duchenne muscular dystrophy model. This line of research seeks to provide innovative delivery strategies for clinical gene therapy, addressing long-standing challenges in vector targeting and immune compatibility.