Highly efficient gene transfer into murine liver achieved by intravenous administration of naked Epstein-Barr virus (EBV)-based plasmid vectors
Highly efficient gene transfer into murine liver achieved by intravenous administration of naked Epstein-Barr virus (EBV)-based plasmid vectors
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DOI：
10.1038/sj.gt.3301551
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发表时间：
2001-10-01
期刊：
影响因子：
5.1
通讯作者：
Mazda, O
中科院分区：
文献类型：
--
作者：
Cui, FD;Kishida, T;Mazda, O
裸质粒DNA(PDNA)注射有望成为病毒和非病毒基因传递系统的替代方法。我们先前已经证明，基于EB病毒(EBV)的含有EBV核抗原1(EBNA 1)基因和ORIP序列的质粒载体能够在各种体外和体内转染系统中实现相当高和持久的表达。以EBV为基础的质粒在高压下通过小鼠尾静脉注射到小鼠体内。标记基因产物在肝脏中大量表达，注射10µg DNA后8~24 h，每克肝脏可表达高达320µg的荧光素酶。转β-半乳糖基因后，70%以上的肝细胞X-半乳糖染色。表达水平显著高于缺乏EBNA-1基因和ORIP的常规pDNA。在转染后第35天，EBV基质粒的表达强度约为传统PDNA基因表达的100倍。EBNA1基因和ORIP都是提高转染率的先决条件。这些结果表明，裸露的EBV载体血管内转染法为体内治疗性基因的肝脏转导提供了一种非常有效、简便的方法。
Naked plasmid DNA (pDNA) injection could become an alternative procedure to viral and nonviral gene delivery systems. We have previously shown that Epstein-Barr virus (EBV)-based plasmid vectors containing the EBV nuclear antigen 1 (EBNA 1) gene and the oriP sequence enable quite high and long-lasting expression in various in vitro and in vivo transfection systems. The EBV-based plasmids were intravenously injected into mice via their tail vein under high pressure. A large amount of the marker gene product was expressed in the liver, as much as 320 mug of luciferase was demonstrated per gram of liver at 8 to 24 h after a single injection with 10 mug of DNA. More than 70% of liver cells stained with X-gal when beta -gal gene was transferred. The expression level was significantly higher than that obtained by conventional pDNA lacking the EBNA 1 gene and oriP. On day 35 after the transfection, the expression from the EBV-based plasmid was approximately 100-fold stronger than the conventional pDNA gene expression. Both the EBNA1 gene and oriP are a prerequisite for the augmentation of the transfection efficiency. These results suggest that the intravascular transfection with naked EBV-based plasmid may provide a quite efficient, simple and convenient means to transduce therapeutic genes in vivo into the liver.