Direct gene transfer into rat liver cells by in vivo electroporation

FEBS Letters

Fujikura, Keiko¹  Shin, Bo-Chul¹  Takata, Kuniaki¹  Suzuki, Takeshi¹  Matsuzaki, Toshiyuki¹ 

[1] Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Showa-machi 3-39-15, Gunma 371-8512, Japan

关键词: Electroporation;    In vivo;    Green fluorescent protein;    Rat liver;    DNA dosage;    Electric pulse;    Local gene therapy;    GFP;    green fluorescent protein;    PBS;    phosphate-buffered saline;    CMV;    cytomegalovirus;   

DOI  :  10.1016/S0014-5793(98)00284-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown