Recombinant adeno-associated viruses (rAAVs) are currently considered the safest and most reliable gene delivery vehicles for human gene therapy. Three serotype capsids, AAV1, AAV2, and AAV9, have been approved for commercial use in patients, but they may not be suitable for all therapeutic contexts. Here, we describe a novel capsid identified in a human clinical sample by high-throughput, long-read sequencing. The capsid, which we have named AAVv66, shares high sequence similarity with AAV2. We demonstrate that compared to AAV2, AAVv66 exhibits enhanced production yields, virion stability, and CNS transduction. Unique structural properties of AAVv66 visualized by cryo-EM at 2.5-Å resolution, suggest that critical residues at the three-fold protrusion and at the interface of the five-fold axis of symmetry likely contribute to the beneficial characteristics of AAVv66. Our findings underscore the potential of AAVv66 as a gene therapy vector.

重组腺相关病毒（rAAV）目前被认为是人类基因治疗最安全、最可靠的基因递送载体。三种血清型衣壳，AAV1、AAV2 和 AAV9，已被批准用于患者的商业用途，但它们可能并不适合所有治疗环境。在这里，我们描述了通过高通量、长读长测序在人类临床样本中鉴定出的一种新型衣壳。我们将衣壳命名为 AAVv66，与 AAV2 具有高度的序列相似性。我们证明，与 AAV2 相比，AAVv66 表现出更高的产量、病毒体稳定性和 CNS 转导。通过冷冻电镜以 2.5-Å 分辨率观察到的 AAVv66 独特的结构特性表明，三重突起和五重对称轴界面处的关键残基可能有助于 AAVv66 的有益特性。我们的研究结果强调了 AAVv66 作为基因治疗载体的潜力。