Chimeric AAV Cap sequences alter gene transduction.
Chimeric AAV Cap sequences alter gene transduction.
复制标题
嵌合 AAV Cap 序列改变基因转导。
DOI：
10.1016/j.virol.2009.01.012
复制
发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Walsh,ChristopherE
中科院分区：
文献类型：
--
作者：
Ward,Peter;Walsh,ChristopherE
重组腺相关病毒（rAAV）是用于基因递送的载体。rAAV存在于显示具有不同转导能力的几种血清型中。血清型的衣壳序列是不同的，表明基因递送的差异在很大程度上是由于衣壳结构。由于可用的血清型在每个细胞的颗粒基础上是细胞系的低效转换器，并且在体内不能有效地转染所需的靶细胞，因此已经进行了许多尝试来通过修饰衣壳来产生更好的载体。一个令人感兴趣的问题是，自然选择是否已经导致，以及基于实验室的选择方法是否会导致具有明确定义的最佳特异性的病毒。在这里，我们使用已知的AAV血清型衣壳序列通过STEP和改组方法创建了多个随机重组的衣壳种类。然后，我们使用选择来鉴定比已知血清型更适合Hep G2细胞的病毒衣壳。然后用其他细胞测试该衣壳，以确定选择是否产生通过选择方法特异性定制的病毒或具有更广泛适用性的病毒。所选择的病毒结果令人惊讶地受到其靶细胞和选择方法的限制。
Recombinant adeno-associated viruses (rAAV) are vectors for gene delivery. rAAVs occur in several serotypes shown to have different transduction capabilities. Capsid sequences of the serotypes are different suggesting that differences in gene delivery are in large part due to capsid structure. Since the available serotypes are inefficient transducers of cell lines on a particle per cell basis and inefficiently transduce desired target cells in vivo there have been numerous attempts to create better vectors by modifying the capsids. A question of interest is whether natural selection has led, and whether laboratory-based selection methods will lead, to viruses with a sharply defined optimal specificity. Here we created multiple randomly recombined capsid species using the known AAV serotype capsid sequences by STEP and shuffling methods. We then used selection to identify a viral capsid better adapted to Hep G2 cells than the known serotypes. This capsid was then tested with other cells to determine whether the selection produced a virus specifically tailored by the selection methodology or one of wider applicability. The selected virus turned out to be surprisingly limited by its target cell and method of selection.