A new adenoviral vector: Replacement of all viral coding sequences with 28 kb of DNA independently expressing both full-length dystrophin and beta-galactosidase
A new adenoviral vector: Replacement of all viral coding sequences with 28 kb of DNA independently expressing both full-length dystrophin and beta-galactosidase
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DOI：
10.1073/pnas.93.12.5731
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发表时间：
1996-06-11
影响因子：
11.1
通讯作者：
Caskey, CT
中科院分区：
文献类型：
--
作者：
Kochanek, S;Clemens, PR;Caskey, CT
腺病毒载体介导的基因转移为许多人类疾病的基因治疗提供了巨大的潜力，然而，由于一些限制，进展缓慢，首先，目前可用的腺病毒载体的插入能力仅限于8 Mb的外源DNA，其次，病毒蛋白在感染细胞中的表达被认为会引发细胞免疫反应，导致炎症，并且只会短暂表达转移的基因。我们报道了一种新的腺病毒载体的发展，它删除了所有的病毒编码序列，因此，大插入被容纳，所有病毒蛋白的表达被消除。该载体系统的首次应用携带了一个双表达盒，包含28.2 kb的非病毒DNA，包括全长鼠肌营养不良蛋白cDNA，由一个大型肌肉特异性启动子和一个lacZ报告基因结构控制。使用该载体，我们证明了这两个基因在原代mdx（肌营养不良蛋白缺乏）肌肉细胞中的独立表达。
Adenoviral vector-mediated gene transfer offers significant potential for gene therapy of many human diseases, However, progress has been slowed by several limitations, First, the insert capacity of currently available adenoviral vectors is limited to 8 Mb of foreign DNA, Second, the expression of viral proteins in infected cells is believed to trigger a cellular immune response that results in inflammation and in only transient expresssion of the transferred gene, We report the development of a new adenoviral vector that has all viral coding sequences removed, Thus, large inserts are accommodated and expression of all viral proteins is eliminated. The first application of this vector system carries a dual expression cassette comprising 28.2 kb of nonviral DNA that includes the full-length murine dystrophin cDNA under control of a large muscle-specific promoter and a lacZ reporter construct. Using this vector, we demonstrate independent expression of both genes in primary mdx (dystrophin-deficient) muscle cells.