DIRECT GENE-TRANSFER INTO RAT-LIVER CELLS BY IN-VIVO ELECTROPORATION

In vivo electro-transfection efficiency and manner of transferred gene expression were investigated by fluorescence microscopic image analys is, Green fluorescent protein (GFP) gene was used as the genetic marke r. Electroporation was carried out on the liver of li re rats by use o f disk electrodes mounted in the tips of tweezers, which were directly pressed onto the surface of a li,er lobe in situ, Electroporation wit h eight electric pulses of 50 ms in duration at 50 V gave a good effic iency 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 tha t the amount of induced gene expression can be controlled. Estimation of the efficiency of electro-gene transfer using the fluorescence of G FP and digital analysis of microscopic images was useful to determine the optimum conditions for local gene therapy in tissues and organs. ( C) 1998 Federation of European Biochemical Societies.