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Journal of Lipid Research, Vol. 41, 1082-1086, July 2000
Copyright © 2000 by Lipid Research, Inc.

Original Article

Adenovirus-mediated gene transfer in primary murine adipocytes

Correspondence to: David A. Bernlohr

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.—Hertzel, A. V., M. A. Sanders, and D. A. Bernlohr. Adenovirus-mediated gene transfer in primary murine adipocytes. J. Lipid Res. 2000. 41: 1082–1086.

Supplementary key words: adipocytes, adenovirus, green fluorescent protein, reporter genes

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