Electroporation-mediated gene transfer that targets glomeruli.
Electroporation has been applied to introducing DNA into several organs; however, gene expression was localized around the injected area. Examined was the efficiency of intrarenal injection of DNA followed by in vivo electroporation, using FITC-labeled oligodeoxynucleotides (FITC-ODN) and plasmid DNA expressing beta-galactosidase or luciferase. FITC-ODN or expression vectors were injected into the left renal artery; thereafter, the left kidney was electroporated between a pair of tweezer-type electrodes. FITC-ODN were transferred into all glomeruli, and transfected cells were identified as mesangial cells. Four d after transfection of the pCAGGS-LacZ gene, beta-galactosidase expression was observed in 75% of glomeruli. To compare the transfection efficacy by electroporation with that by the hemagglutinating virus of Japan (HVJ) liposome method, a luciferase reporter gene, pActLuc, was transferred into glomeruli by either electroporation or the HVJ liposome method. On day 4, electroporation resulted in higher glomerular luciferase activity than did the HVJ liposome method. We also observed that co-transfection of pcEBNA, an expression vector for Epstein-Barr virus nuclear antigen, and poriP-cLuc, oriP-harboring vector, resulted in an eightfold higher luciferase gene expression than simple poriP-cLUC: No histologic damages were seen in glomeruli or tubular epithelial cells. In conclusion, gene transfer into renal artery followed by electroporation was an effective and simple strategy for gene transfer that targets glomerular mesangial cells.
Tsujie
Michiko
M
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Isaka
Yoshitaka
Y
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Nakamura
Hiroyuki
H
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Imai
Enyu
E
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Hori
Masatsugu
M
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D016650:Fluorescein-5-isothiocyanate; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005920:Glomerular Mesangium / Q000378:metabolism; D007678:Kidney Glomerulus / Q000378:metabolism*; D007763:Lac Operon; D008156:Luciferases / Q000235:genetics; D008297:Male; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012077:Renal Artery; D014162:Transfection
D017931:DNA Primers; D019309:Epstein-Barr Virus Nuclear Antigens; D009838:Oligodeoxyribonucleotides; D008156:Luciferases; D016650:Fluorescein-5-isothiocyanate