Adeno-associated virus (AAV) is a non-enveloped virus that has emerged as a promising gene delivery vector due to its safety and efficacy in both preclinical and clinical settings. This review highlights the key aspects of using AAV as a gene therapy vector, including its versatility, safety profile, and potential for a wide range of clinical applications. AAV vectors can be engineered to deliver DNA to target cells, and their unique biological and biophysical properties make them suitable for treating various diseases. The review covers the design considerations for AAV vectors, such as packaging size, promoter selection, and transgene optimization. It also discusses the selection and optimization of AAV capsids to enhance transduction efficiency and reduce immunogenicity. The manufacturing processes for clinical use, including transient transfection platforms and producer cell lines, are described, along with the challenges and advancements in purification and downstream processing. Finally, the review explores different delivery strategies for AAV-based therapies, such as systemic, intramuscular, central nervous system, cardiac, and pulmonary delivery, and their potential applications in treating various diseases.Adeno-associated virus (AAV) is a non-enveloped virus that has emerged as a promising gene delivery vector due to its safety and efficacy in both preclinical and clinical settings. This review highlights the key aspects of using AAV as a gene therapy vector, including its versatility, safety profile, and potential for a wide range of clinical applications. AAV vectors can be engineered to deliver DNA to target cells, and their unique biological and biophysical properties make them suitable for treating various diseases. The review covers the design considerations for AAV vectors, such as packaging size, promoter selection, and transgene optimization. It also discusses the selection and optimization of AAV capsids to enhance transduction efficiency and reduce immunogenicity. The manufacturing processes for clinical use, including transient transfection platforms and producer cell lines, are described, along with the challenges and advancements in purification and downstream processing. Finally, the review explores different delivery strategies for AAV-based therapies, such as systemic, intramuscular, central nervous system, cardiac, and pulmonary delivery, and their potential applications in treating various diseases.