Intracellular Gene Transfer in Rats by Tail Vein Injection of Plasmid DNA
Intracellular Gene Transfer in Rats by Tail Vein Injection of Plasmid DNA
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DOI：
10.1208/s12248-010-9231-z
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发表时间：
2010-09
期刊：
影响因子：
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通讯作者：
Tian Zhou;Kenya Kamimura;Guisheng Zhang;Dexi Liu
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文献类型：
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作者：
Tian Zhou;Kenya Kamimura;Guisheng Zhang;Dexi Liu
本研究探讨了多种因素对质粒DNA尾静脉注射大鼠基因转染效率的影响。我们测量了报告基因在内脏器官中的表达水平，包括肺、心脏、脾、肾和肝，作为注射体积、注射时间和DNA剂量的函数。还检查了转染动物中报告基因表达的持续性。我们证明，质粒递送到大鼠的尾静脉是有效的，只要注射的DNA溶液的体积被调整到7-8%的体重，注射时间小于10秒。除了在注射期间和注射后不久血清丙氨酸氨基转移酶浓度的短期升高和心脏功能的短暂不规则性外，该程序耐受性良好。来自转染动物的肝脏的Lac Z染色显示约5-10%的阳性细胞。使用携带由鸡β肌动蛋白基因启动子驱动的人α 1抗胰蛋白酶基因cDNA的质粒和CMV增强子在动物中进行转基因表达的持久性测试显示，转基因产物在注射后1天达到峰值水平，随后随着时间的推移逐渐下降。在第一次注射后第38天进行第二次注射，重新获得峰值水平。结果表明，尾静脉注射是一种有效的质粒DNA导入大鼠肝细胞的方法。我们相信，这一程序将是非常有用的基因功能研究的背景下，整个动物在大鼠。
In this study, we examined the effect of various factors on gene delivery efficiency of tail vein injection of plasmid DNA into rats. We measured the level of reporter gene expression in the internal organs including the lung, heart, spleen, kidney, and liver as function of injection volume, injection time, and DNA dose. Persistency of reporter gene expression in transfected animals was also examined. We demonstrated that plasmid delivery to rats by the tail vein is effective as long as the volume of injected DNA solution is adjusted to 7–8% of body weight with an injection time of less than 10 s. With the exception of a short-term increase in serum concentration of alanine aminotransferase and transient irregularity in cardiac function during and soon after the injection, the procedure is well tolerated. Lac Z staining of the liver from transfected animals showed approximately 5–10% positive cells. Persistency test for transgene expression in animals using plasmid carrying cDNA of human alpha 1 antitrypsin gene driven by chicken beta actin gene promoter with CMV enhancers showed peak level of transgene product 1 day after the injection followed by a gradual decline with time. Peak level was regained by a second injection performed on day 38 after the first injection. These results show that tail vein injection is an effective means for introducing plasmid DNA into liver cells in rats. We believe that this procedure will be extremely useful for gene function studies in the context of whole animal in rats.