Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA
Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA
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DOI：
10.1038/sj.gt.3300947
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发表时间：
1999-07-01
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：
Liu, F;Song, YK;Liu, D
开发能够在动物体内有效表达外源基因的方法，将为基因功能研究、疾病治疗和获得基因产品提供工具。因此，我们开发了一种基于流体动力学的程序，通过系统给药质粒DNA在小鼠中表达转基因。利用荧光素酶和β -半乳糖苷酶的cDNA作为报告基因，我们证明了通过失败静脉快速注射大量的DMA溶液可以实现有效的基因转移和表达。在表达该基因的器官中，肝脏的基因表达量最高。通过向小鼠尾静脉注射5 μ g质粒DNA，每克肝脏可获得高达45 μ g的荧光素酶蛋白。使用β -半乳糖苷酶基因作为报告基因的组织化学分析显示，大约40%的肝细胞表达转基因。时间响应曲线显示，在注射后约8 h，肝脏中转基因表达水平达到峰值，随后逐渐下降。反复注射质粒DNA可恢复基因表达的峰值水平。这些结果表明，我们已经开发出一种简单、方便、高效的方法，可以作为通过基因转移研究基因功能、基因调控和分子病理生理以及在动物体内表达蛋白质的有效手段。
Development of methods that allow an efficient expression of exogenous genes in animals would provide tools for gene function studies, treatment of diseases and for obtaining gene products. Therefore, we have developed a hydrodynamics-based procedure for expressing transgenes in mice by systemic administration of plasmid DNA. Using cDNA of luciferase and beta-galactosidase as a reporter gene, we demonstrated that an efficient gene transfer and expression can be achieved by a rapid injection of a large volume of DMA solution into animals via the fail vein. Among the organs expressing the transgene, the liver showed the highest level of gene expression. AS high as 45 mu g of luciferase protein per gram of liver can be achieved by a single tail vein injection of 5 mu g of plasmid DNA into a mouse. Histochemical analysis using beta-galactosidase gene as a reporter reveals that approximately 40% of hepatocytes express the transgene. The time-response curve shows that the level of transgene expression in the liver reaches the peak level in approximately 8 h after injection and decreases thereafter. The peak level of gene expression can be regained by repeated injection of plasmid DNA. These results suggest that a simple, convenient and efficient method has been developed and which can be used as an effective means for studying gene function, gene regulation and molecular pathophysiology through gene-transfer, as well as for expressing proteins in animals.