Development of gene transfer by in vivo electroporation to the vascular wall

我们开发了一种用质粒DNA在体内电穿孔进行动脉基因转移的非病毒方法。电穿孔是通过对细胞或组织施加短时间、高强度的电场脉冲，在细胞膜上形成纳米化瞬态孔的物理过程。有效电穿孔的最重要参数是：刺激电压、脉冲持续时间、电流、质粒浓度和电极形状。结果：30电压、20 msec脉冲穿孔使血管壁基因表达显著增加。质粒DNA浓度为400μ g/ml时效果最佳。通过体内电穿孔转移的血管基因的咬合集中在电极边缘周围。血管壁模拟模型和光学测绘实验也证实了这一结论。我们比较了单板电极（5^*1mm板）和多板电极（两个平行板）在血管壁上的基因表达。多片电极的基因表达优于单片电极。综上所述，我们开发了一种利用电穿孔进行基因转移的方法，这种方法被证明比使用裸DNA更安全，更有效。然后，将来就有可能处理基因表达的位点或数量。

We developed a nonviral method for arterial gene transfer by in vivo electroporation with plasmid DNA. Electroporation is the physical process of including nanometreaized transient pores in a cell membrane by the application of short duration, high intensity electric field pulses to cells or tissues. The most important parameters for effective electroporation are: the stimulation voltage, the pulse duration, the electric current, plasmid concentration, and the shape of an electrode.Result: Electroporation with 30 voltage and pulse duration of 20 msec resulted in a marked increase of gene expression in vessel wall. The concentration of 400μ g/ml plasmid DNA is most optimal. The Bite of the vessel gene-transferred by in vivo electroporation is focused around the edge of the electrode. The same findings are demonstrated by the simulation model of vessel wall and the experiment of optical mapping. We compare with gene expression in vessel wall between with the monoplate electrode, which is one 5^*1mm plate, and the multiplate electrode, which is two plates in parallel. The gene expression using muitiplate electrode is superior to the one of monoplate electrode.In summary, we developed a method for gene transfer using electroporation, an approach which proved to the safe and more efficient than that using naked DNA. And then it will be possible that the site or the mount of gene expression is handled in the future.