Impaired intracellular trafficking of adeno-associated virus type 2 vectors limits efficient transduction of murine fibroblasts
Impaired intracellular trafficking of adeno-associated virus type 2 vectors limits efficient transduction of murine fibroblasts
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DOI：
10.1128/jvi.74.2.992-996.2000
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发表时间：
2000-01-01
影响因子：
5.4
通讯作者：
Srivastava, A
中科院分区：
文献类型：
--
作者：
Hansen, J;Qing, K;Srivastava, A
尽管2型腺相关病毒（AAV）作为更常用的基于逆转录病毒和腺病毒的载体用于人基因治疗的潜在有用的替代物而受到关注，但通过AAV载体的有效基因转移和转基因表达需要克服以下两个障碍。首先，靶细胞必须表达AAV感染的受体和辅助受体，其次，细胞必须允许病毒第二链DNA合成。我们现在描述的第三个障碍，受损的细胞内运输的AAV的细胞核，这导致在缺乏转基因表达的小鼠成纤维细胞，表达的AAV受体和辅助受体，并允许病毒的第二链DNA合成。我们记录了AAV载体有效地结合并成功进入NIH 3T3细胞，但在这些细胞中进入细胞核的运输显著受损。相比之下，已知通过AAV载体有效转导的细胞中病毒运输至细胞核是快速且有效的。在AAV介导的基因转移中的另一个障碍的证明对这些载体在人类基因治疗中的最佳使用具有影响。
Although adeno-associated virus type 2 (AAV) has gained attention as a potentially useful alternative to the more commonly used retrovirus- and adenovirus based vectors for human gene therapy, efficient gene transfer and transgene expression by AAV vectors require that the following two obstacles be overcome. First, the target cell must express the receptor and the coreceptor for AAV infection, and second, the cell must allow for viral second-strand DNA synthesis. We now describe a third obstacle, impaired intracellular trafficking of AAV to the nucleus, which results in the lack of transgene expression in murine fibroblasts which do express the AAV receptor and the coreceptor and which are permissive for viral second-strand DNA synthesis. We document that AAV vectors bind efficiently and gain entry successfully into NIH 3T3 cells, but trafficking into the nucleus is significantly impaired in these cells. In contrast, viral trafficking to the nucleus in cells known to be efficiently transduced by AAV vectors is both rapid and efficient. The demonstration of yet another obstacle in AAV-mediated gene transfer has implications for the optimal use of these vectors in human gene therapy.