Adeno-associated virus (AAV) has emerged as a pivotal delivery tool in clinical gene therapy owing to its minimal pathogenicity and ability to establish long-term gene expression in different tissues. Recombinant AAV (rAAV) has been engineered for enhanced specificity and developed as a tool for treating various diseases. However, as rAAV is being more widely used as a therapy, the increased demand has created challenges for the existing manufacturing methods. Seven rAAV-based gene therapy products have received regulatory approval, but there continue to be concerns about safely using high-dose viral therapies in humans, including immune responses and adverse effects such as genotoxicity, hepatotoxicity, thrombotic microangiopathy, and neurotoxicity. In this review, we explore AAV biology with an emphasis on current vector engineering strategies and manufacturing technologies. We discuss how rAAVs are being employed in ongoing clinical trials for ocular, neurological, metabolic, hematological, neuromuscular, and cardiovascular diseases as well as cancers. We outline immune responses triggered by rAAV, address associated side effects, and discuss strategies to mitigate these reactions. We hope that discussing recent advancements and current challenges in the field will be a helpful guide for researchers and clinicians navigating the ever-evolving landscape of rAAV-based gene therapy.

腺相关病毒（AAV）由于其致病性极小且能够在不同组织中建立长期基因表达，已成为临床基因治疗中的关键递送工具。重组 AAV (rAAV) 已被设计用于增强特异性，并被开发为治疗各种疾病的工具。然而，随着rAAV作为一种治疗方法的应用越来越广泛，需求的增加给现有的制造方法带来了挑战。七种基于 rAAV 的基因治疗产品已获得监管部门批准，但人们仍然担心在人类中安全使用高剂量病毒疗法，包括免疫反应和遗传毒性、肝毒性、血栓性微血管病和神经毒性等不良反应。在这篇综述中，我们探讨了 AAV 生物学，重点是当前的载体工程策略和制造技术。我们讨论了 rAAV 如何用于正在进行的眼科、神经、代谢、血液、神经肌肉、心血管疾病以及癌症的临床试验。我们概述了 rAAV 触发的免疫反应，解决相关的副作用，并讨论减轻这些反应的策略。我们希望讨论该领域的最新进展和当前挑战将为研究人员和临床医生应对基于 rAAV 的基因治疗不断发展的前景提供有用的指导。