Long-term expression of human alpha 1-antitrypsin gene in mouse liver achieved by intravenous administration of plasmid DNA using a hydrodynamics-based procedure
Long-term expression of human alpha 1-antitrypsin gene in mouse liver achieved by intravenous administration of plasmid DNA using a hydrodynamics-based procedure
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DOI：
10.1038/sj.gt.3301229
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发表时间：
2000-08-01
期刊：
影响因子：
5.1
通讯作者：
Liu, D
中科院分区：
文献类型：
--
作者：
Zhang, G;Song, YK;Liu, D
肝脏是基因转移的重要靶器官，因为它具有合成血清蛋白的巨大能力，并且与许多遗传性和获得性疾病有关。此前，我们和其他人已经证明，使用基于流体动力学的程序静脉注射质粒 DNA 可以实现体内肝细胞的有效基因转移。在这项研究中，我们系统地表征了编码分泌蛋白的转基因在小鼠中的表达。我们以人α1-抗胰蛋白酶（hAAT）基因为报告基因，通过尾静脉注射10-50μg质粒DNA到小鼠体内，hAAT的血清水平可高达0.5mg/ml。 DNA注射后1天，血清hAAT达到峰值，然后在接下来的2至4周内下降至2-5μg/ml，该水平持续至少6个月。对提取的 DNA 进行 Southern 分析，并对肝脏 RNA 进行 RT-PCR 分析，结果表明 hAAT 基因处于活跃状态，并在 6 个月后以游离形式存在。这些结果表明，基于流体动力学的转染程序为在整个动物中筛选用于治疗目的的基因提供了有价值的工具。
The liver is an important target organ for gene transfer due to its large capacity for synthesizing serum proteins and its involvement in numerous genetic and acquired diseases. Previously, we and others have shown that an efficient gene transfer to liver cells in vivo can be achieved by an intravenous injection of plasmid DNA using a hydrodynamics-based procedure. In this study, we systematically characterized the expression of transgene encoding a secretory protein in mouse. Using human alpha 1-antitrypsin (hAAT) gene as a reporter, we demonstrate that the serum level of hAAT can reach as high as 0.5 mg/ml by a simple tail vein injection of 10-50 mu g plasmid DNA into a mouse. The serum hAAT reaches the peak level 1 day after DNA injection and then declines during the following 2 to 4 weeks to 2-5 mu g/ml, a level which persists for at least 6 months. Southern analysis of extracted DNA and RT-PCR analysis of RNA from the liver reveal that hAAT gene is active and present as episomal form after 6 months. These results suggest that the hydrodynamics-based transfection procedure provides a valuable tool for screening genes for therapeutic purposes in whole animals.