Replacing or editing disease-causing mutations holds great promise for treating many human diseases. Yet, delivering therapeutic genetic modifiers to specific cells in vivo has been challenging, particularly in large, anatomically distributed tissues such as skeletal muscle. Here, we establish an in vivo strategy to evolve and stringently select capsid variants of adeno-associated viruses (AAVs) that enable potent delivery to desired tissues. Using this method, we identify a class of RGD motif-containing capsids that transduces muscle with superior efficiency and selectivity after intravenous injection in mice and non-human primates. We demonstrate substantially enhanced potency and therapeutic efficacy of these engineered vectors compared to naturally occurring AAV capsids in two mouse models of genetic muscle disease. The top capsid variants from our selection approach show conserved potency for delivery across a variety of inbred mouse strains, and in cynomolgus macaques and human primary myotubes, with transduction dependent on target cell expressed integrin heterodimers.

替换或编辑致病突变为治疗许多人类疾病带来了巨大希望。然而，向体内特定细胞提供治疗性遗传修饰剂一直具有挑战性，特别是在骨骼肌等大型解剖学分布组织中。在这里，我们建立了一种体内策略来进化并严格选择腺相关病毒（AAV）的衣壳变体，从而能够有效地将其递送到所需的组织。使用这种方法，我们鉴定了一类含有 RGD 基序的衣壳，在小鼠和非人类灵长类动物静脉注射后，其能够以优异的效率和选择性转导肌肉。我们在两种遗传性肌肉疾病小鼠模型中证明，与天然存在的 AAV 衣壳相比，这些工程载体的效力和治疗功效显着增强。我们的选择方法中的顶级衣壳变体显示出在各种近交系小鼠品系以及食蟹猴和人类原代肌管中传递的保守效力，转导依赖于靶细胞表达的整合素异二聚体。