Multiparametric domain insertional profiling of Adeno-Associated Virus VP1.
Multiparametric domain insertional profiling of Adeno-Associated Virus VP1.
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腺相关病毒 VP1 的多参数域插入分析。
DOI：
10.1101/2023.04.19.537549
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Schmidt,Daniel
中科院分区：
文献类型：
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作者：
Hoffmann,MareikeD;Zdechlik,AlinaC;He,Yungui;Nedrud,David;Aslanidi,George;Gordon,Wendy;Schmidt,Daniel
腺相关病毒（AAV）的一些进化特性，如广泛的嗜性和免疫原性，是基于AAV的基因治疗的障碍。大多数重新设计这些特性的努力都集中在AAV的3倍突起和衣壳蛋白末端附近的可变区。为了全面调查AAV衣壳的可工程化热点，我们在将六个结构化蛋白质结构域插入整个AAV-DJ衣壳蛋白VP 1后确定了多种AAV适应性表型。这是迄今为止最大和最全面的AAV结构域插入数据集。我们的数据揭示了AAV衣壳适应大结构域插入的令人惊讶的稳健性。插入允许性强烈依赖于插入位置、结构域类型和测量的适应性表型，其聚集成连续的结构单元，我们可以将其与AAV组装、稳定性和感染性中的不同作用联系起来。我们还鉴定了促进结合支架的共价连接的AAV的工程化热点，这可能代表了重新定向AAV向性的替代方法。
Several evolved properties of adeno-associated virus (AAV), such as broad tropism and immunogenicity in humans, are barriers to AAV-based gene therapy. Most efforts to re-engineer these properties have focused on variable regions near AAV's 3-fold protrusions and capsid protein termini. To comprehensively survey AAV capsids for engineerable hotspots, we determined multiple AAV fitness phenotypes upon insertion of six structured protein domains into the entire AAV-DJ capsid protein VP1. This is the largest and most comprehensive AAV domain insertion dataset to date. Our data revealed a surprising robustness of AAV capsids to accommodate large domain insertions. Insertion permissibility depended strongly on insertion position, domain type, and measured fitness phenotype, which clustered into contiguous structural units that we could link to distinct roles in AAV assembly, stability, and infectivity. We also identified engineerable hotspots of AAV that facilitate the covalent attachment of binding scaffolds, which may represent an alternative approach to re-direct AAV tropism.