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

Z. Tao, D. Herndon, H. Hawkins, T. Wood, R. Perez-Polo

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

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 β-galactosidase cDNA/CMV construct using cholesterol containing cationic liposomes and measured β-galactosidase and IGF-I mRNA and protein. In vivo, we subcutaneously injected 3-month-old male Sprague-Dawley rats with IGF-I cDNA and β-galactosidase cDNA into rat skin. After IGF-I and β-galactosidase were cotransfected into PCI2 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 β-galactosidase activity confirmed that transfections resulted in a restricted expression area.

Original languageEnglish (US)
Pages (from-to)41-55
Number of pages15
JournalBiochemical Genetics
Volume38
Issue number1-2
DOIs
StatePublished - 2000

Keywords

  • Gene transfer
  • IGF-I
  • Liposome
  • Rat
  • Wound healing

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Biochemistry
  • Molecular Biology
  • Genetics

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