Insulin-like growth factor-I cDNA gene transfer in vitro and in vivo

Biochem Genet. 2000 Feb;38(1-2):41-55. doi: 10.1023/a:1001880220039.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Complementary / administration & dosage*
  • Drug Carriers
  • Gene Expression
  • Genes, Reporter
  • Injections, Intradermal
  • Insulin-Like Growth Factor I / genetics*
  • Male
  • PC12 Cells
  • Rats
  • Skin
  • Transfection*
  • beta-Galactosidase / genetics

Substances

  • DNA, Complementary
  • Drug Carriers
  • Insulin-Like Growth Factor I
  • beta-Galactosidase