Gene therapy, which treats genetic diseases by fixing defective genes, has gained significant attention. Viral vectors show great potential for gene delivery but face limitations like low transduction efficiency and poor targeting. Loading viral vectors onto tissue engineered scaffolds presents a promising strategy to address these challenges, but their widespread application remains limited by challenges like vector stability, biomaterial selection, and high manufacturing costs. Adeno-associated virus (AAV), recognized for its safety, high efficiency, and low immunogenicity, was employed as a model virus. In this study, AAV was encapsulated within electrospun fibers (AAV/PCL-PEO@Co-ES) composed of polycaprolactone (PCL) and polyethylene oxide (PEO) via coaxial electrospinning, ensuring effective AAV protection and controlled release. The physicochemical characterization results indicated that the scaffold exhibited excellent mechanical properties (tensile strength: 3.22 ± 0.48 MPa) and wettability (WCA: 67.90 ± 8.45°). In vitro release and cell transduction assays demonstrated that the AAV-loaded scaffold effectively control viral vector release and transduction. Furthermore, the in vitro cell and in vivo animal experiments suggested that the AAV-loaded scaffolds exhibit excellent biocompatibility and efficient viral vector delivery capability. Hence, our research not only enhances the storage and delivery of viral vectors but also provides innovative solutions for viral vector delivery strategies.