Site-Specific Post-translational Surface Modification of Adeno-Associated Virus Vectors Using Leucine Zippers.
Site-Specific Post-translational Surface Modification of Adeno-Associated Virus Vectors Using Leucine Zippers.
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DOI：
10.1021/acssynbio.9b00341
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发表时间：
2020-02
影响因子：
4.7
通讯作者：
Nicole N. Thadani;Joanna Yang;Buhle Moyo;Ciaran M. Lee;Maria Y. Chen;Gang Bao;J. Suh
中科院分区：
文献类型：
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作者：
Nicole N. Thadani;Joanna Yang;Buhle Moyo;Ciaran M. Lee;Maria Y. Chen;Gang Bao;J. Suh
腺相关病毒（AAV）作为基因治疗载体受到广泛青睐，在国际上进行了200多项临床试验。为了改善靶向递送，已经探索了多种遗传衣壳修饰，例如将靶向蛋白/肽插入衣壳壳中，并取得了一些成功，但较大的插入通常对衣壳形成和基因递送具有不可预测的有害影响。在这里，我们展示了一个模块化平台，用于将外源肽和蛋白质整合到AAV衣壳上，同时保留载体功能。我们用亮氨酸拉链卷曲螺旋结合基序装饰AAV衣壳，这些基序表现出特异性非共价异源二聚化。AAV衣壳使用这种方法成功地展示了六组氨酸标记的肽，如通过镍柱亲和性所证明的。这种蛋白质展示平台可以促进生物部分在AAV表面上的掺入，从而扩大载体增强和工程化的可能性。
Adeno-associated virus (AAV) is widely favored as a gene therapy vector, tested in over 200 clinical trials internationally. To improve targeted delivery a variety of genetic capsid modifications, such as insertion of targeting proteins/peptides into the capsid shell, have been explored with some success but larger insertions often have unpredictable deleterious impacts on capsid formation and gene delivery. Here, we demonstrate a modular platform for the integration of exogenous peptides and proteins onto the AAV capsid post-translationally while preserving vector functionality. We decorated the AAV capsid with leucine-zipper coiled-coil binding motifs that exhibit specific noncovalent heterodimerization. AAV capsids successfully display hexahistidine tagged-peptides using this approach, as demonstrated through nickel column affinity. This protein display platform may facilitate the incorporation of biological moieties on the AAV surface, expanding possibilities for vector enhancement and engineering.