Insulin-like growth factor-I cDNA gene transfer in vitro and in vivo.
Our hypothesis is that gene transfer of an IGF-I CMV-cDNA with cholesterol containing cationic liposomes is an efficient tool for transient transfection of growth factors in vitro and in vivo. In vitro, we transiently cotransfected IGF-I cDNA with a CMV construct and a Lac Z beta-galactosidase cDNA/CMV construct using cholesterol containing cationic liposomes and measured beta-galactosidase and IGF-I mRNA and protein. In vivo, we subcutaneously injected 3-month-old male Sprague-Dawley rats with IGF-I cDNA and beta-galactosidase cDNA into rat skin. After IGF-I and beta-galactosidase were cotransfected into PC12 cells, Northern blot analysis showed that the peak time of IGF-I expression was 2 days for mRNA and 5 days for protein. In vivo, a cDNA/liposome ratio of 1:2 was most effective. IGF-I protein expression in IGF-I-transfected skin resulted in significant transfection from day 5 to day 7. In situ determination of beta-galactosidase activity confirmed that transfections resulted in a restricted expression area.
Tao
Z
Z
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch at Galveston 77555, USA.
Herndon
D
D
Hawkins
H
H
Wood
T
T
Perez-Polo
R
R
D000818:Animals; D018076:DNA, Complementary / Q000008:administration & dosage*; D004337:Drug Carriers; D015870:Gene Expression; D017930:Genes, Reporter; D007271:Injections, Intradermal; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D008297:Male; D016716:PC12 Cells; D051381:Rats; D012867:Skin; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D018076:DNA, Complementary; D004337:Drug Carriers; D007334:Insulin-Like Growth Factor I; D001616:beta-Galactosidase