Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis
Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis
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DOI：
10.1128/jvi.70.1.520-532.1996
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发表时间：
1996-01-01
影响因子：
5.4
通讯作者：
Wilson, JM
中科院分区：
文献类型：
--
作者：
Fisher, KJ;Gao, GP;Wilson, JM
腺相关病毒是一种整合DNA的细小病毒，有可能成为体细胞基因治疗的重要载体。然而，一个潜在的障碍是重组腺相关病毒（rAAV）载体的低转导效率。我们在这份报告中表明，腺病毒显着增强rAAV在体外转导的方式是依赖于早期区域1和4的表达（分别为E1和E4）基因，并且与rAAV基因组的双链复制形式的出现成正比。在E1不存在的情况下，来自E4的开放阅读框6蛋白的表达实现了类似但减弱的效果，通过使用肝和肺定向基因治疗的鼠模型，在体内类似地证明了腺病毒El和E4对于rAAV基因转移的辅助活性。我们的数据表明单链rAAV基因组向双链体中间体的转化限制了基因治疗的转导和有用性。
Adeno-associated virus is an integrating DNA parvovirus with the potential to be an important vehicle for somatic gene therapy. A potential barrier, however, is the low transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors. We show in this report that adenovirus dramatically enhances rAAV transduction in vitro in a way that is dependent on expression of early region 1 and 4 (E1 and E4, respectively) genes and directly proportional to the appearance of double-stranded replicative forms of the rAAV genome, Expression of the open reading frame 6 protein from E4 in the absence of El accomplished a similar but attenuated effect, The helper activity of adenovirus El and E4 for rAAV gene transfer was similarly demonstrated in vivo by using murine models of liver- and lung-directed gene therapy. Our data indicate that conversion of a single-stranded rAAV genome to a duplex intermediate limits transduction and usefulness for gene therapy.