Adeno-associated virus (AAV) has emerged as a viral gene delivery vector that is safe in humans, able to infect both dividing and arrested cells and drive long-term expression (>6 months). Unfortunately, the naturally evolved properties of many AAV serotypes-including low cell type specificity and largely overlapping tropism-are mismatched to applications that require cell type-specific infection, such as neural circuit mapping or precision gene therapy. A variety of approaches to redirect AAV tropism exist, but there is still the need for a universal solution for directing AAV tropism toward user-defined cellular receptors that does not require extensive case-by-case optimization and works with readily available components. Here, we report AAV engineering approaches that enable programmable receptor-mediated gene delivery. First, we genetically encode small targeting scaffolds into a variable region of an AAV capsid and show that this redirects tropism toward the receptor recognized by these targeting scaffolds and also renders this AAV variant resistant to neutralizing antibodies present in nonhuman primate serum. We then simplify retargeting of tropism by engineering the same variable loop to encode a HUH tag, which forms a covalent bond to single-stranded DNA oligos conjugated to store-bought antibodies. We demonstrate that retargeting this HUH-AAVs toward different receptors is as simple as "arming" a premade noninfective AAV template with a different antibody in a conjugation process that uses widely available reagents and requires no optimization or extensive purification. Composite antibody-AAV nanoparticles structurally separate tropism and payload encapsulation, allowing each to be engineered independently.

中文翻译：

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用于受体介导的基因传递的腺相关病毒-抗体复合物的可编程组装。

腺相关病毒（AAV）已作为一种对人类安全的病毒基因传递载体出现，能够感染分裂和停滞的细胞并驱动长期表达（> 6个月）。不幸的是，许多AAV血清型的自然进化特性（包括低细胞类型特异性和很大程度上重叠的嗜性）与需要细胞类型特异性感染的应用（例如神经回路作图或精密基因治疗）不匹配。存在多种重定向AAV向性的方法，但是仍然需要用于将AAV向性引向用户定义的细胞受体的通用解决方案，该解决方案不需要进行逐案优化，并且可以使用容易获得的组件来工作。在这里，我们报告了AAV工程方法，使可编程受体介导的基因传递成为可能。第一的，我们将小型靶向支架遗传编码到AAV衣壳的可变区中，并显示这将向性重定向到这些靶向支架识别的受体，并使该AAV变体对中和非人类灵长类动物血清中的抗体具有抗性。然后，我们通过工程化相同的可变环来编码HUH标签，从而简化了向向性的重定靶标，该标签形成了共价键，该共价键与偶联至存储购买抗体的单链DNA寡核苷酸形成共价键。我们证明，将这种HUH-AAVs重定向到不同的受体就像在使用广泛可用的试剂进行偶联的过程中，用不同的抗体“武装”预制的非感染性AAV模板一样简单，不需要优化或大量纯化。