Suzuki T, Shin B C, Fujikura K, Matsuzaki T, Takata K
Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, Japan.
FEBS Lett. 1998 Apr 3;425(3):436-40. doi: 10.1016/s0014-5793(98)00284-1.
In vivo electro-transfection efficiency and manner of transferred gene expression were investigated by fluorescence microscopic image analysis. Green fluorescent protein (GFP) gene was used as the genetic marker. Electroporation was carried out on the liver of live rats by use of disk electrodes mounted in the tips of tweezers, which were directly pressed onto the surface of a liver lobe in situ. Electroporation with eight electric pulses of 50 ms in duration at 50 V gave a good efficiency of transfection as judged by the induced GFP expression. Bright fluorescence of GFP appeared as dots, which were scattered around the area damaged by electroporation. The transfection efficiency increased as the amount of injected DNA was increased. The results indicate that the amount of induced gene expression can be controlled. Estimation of the efficiency of electro-gene transfer using the fluorescence of GFP and digital analysis of microscopic images was useful to determine the optimum conditions for local gene therapy in tissues and organs.
通过荧光显微镜图像分析研究了体内电转染效率和转移基因的表达方式。使用绿色荧光蛋白（GFP）基因作为遗传标记。利用安装在镊子尖端的圆盘电极对活大鼠的肝脏进行电穿孔，将电极直接原位按压在肝叶表面。以50V施加八个持续时间为50ms的电脉冲进行电穿孔，根据诱导的GFP表达判断，转染效率良好。GFP的明亮荧光呈点状出现，散布在电穿孔损伤区域周围。转染效率随注射DNA量的增加而提高。结果表明，可以控制诱导基因表达的量。利用GFP荧光和显微镜图像的数字分析来估计电基因转移效率，有助于确定组织和器官局部基因治疗的最佳条件。
