Adenovirus-mediated gene transfer in primary murine adipocytes

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

The transfer of genes into primary murine adipocytes using an adenovirus system has been developed. A recombinant adenovirus was constructed (expressing green fluorescent protein [GFP] under the control of the strong cytomegalovirus [CMV] promoter and a luciferase reporter gene under the control of the weak adipocyte promoter keratinocyte lipid-binding protein [KLBP/FABP5]) and incubated with primary adipocytes from C57BL/6J mice. Analysis of infected cells by confocal microscopy detected GFP expression in both the cytoplasm and nucleus of adipocytes with a 64% efficiency of infection. To demonstrate the applicability of this method in the study of gene regulation, adenovirus-infected adipocytes exhibited significant levels of luciferase activity even from a weak promoter. TPA treatment of infected adipocytes increased luciferase activity, consistent with previous studies indicating that the KLBP/FABP5 gene is up-regulated by phorbol esters. These results provide an efficient, convenient, and sensitive method to transiently infect primary murine adipocytes, facilitating protein expression or permitting analysis of reporter gene activity from both viral and endogenous promoters.

Original languageEnglish (US)
Pages (from-to)1082-1086
Number of pages5
JournalJournal of lipid research
Volume41
Issue number7
StatePublished - Jul 2000

Keywords

  • Adenovirus
  • Adipocytes
  • Green fluorescent protein
  • Reporter genes

Fingerprint Dive into the research topics of 'Adenovirus-mediated gene transfer in primary murine adipocytes'. Together they form a unique fingerprint.

Cite this