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KMID : 1161519980020020249
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Animal Cells and Systems
1998 Volume.2 No. 2 p.249 ~ p.258
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Construction and characterization of novel expression vectors for genetic adipose tissue ablation
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Ko Duck-Sung
Choi Woong-Hwan
Kim Chul-Geun
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Abstract
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Obesity, one of the most common metabolic diseases in industrial countries is characterized by an increase in the number or size of adipocytes. In an effort to create transgenic mouse models for the study of obesity, we developed a novel technique in which adipose tissue can be ablated genetically at will, at any specific developmental stage and/or physiological condition, by the treatment of ganciclovir. We made a series of adipocyte?specific expression vectors using minimal regulatory regions of brown adipocyte?specific uncoupling protein (UCP?1) gene and adipocyte?specific aP2 gene, and then analyzed their expression characteristics in cultured cell lines. When both constructs pUCP?LacZ and paP2?LacZ were trans?fected transiently into differentiating 3T3?L1 (pre?white adipocytes) and HIB?1B (pre?brown adipocytes) cell lines in vitro and then monitored by X?gal staining of cells, these regulatory regions were sufficient to show proper differentiation stage?specific expression in adipocytes. To confirm that adipocytes expressing HSV?TK controlled by these minimal regulatory elements are sufficient to kill themselves with ganciclovir treatment, pUCP?TK and paP2?TK expression constructs were transfected stably into HIB?1B and 3T3?L1 cells, respectively, and their ganciclovir sensitivities were tested during in vitro differentiation of cells. As expected, more than 80% of cells were dead by the 7th day of treatment with ganciclovir, while negative control cells were not affected at all. The data suggest that the constructed vectors are suitable for obtaining novel obese transgenic models based on a conditional genetic tissue ablation method
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KEYWORD
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Genetic tissue ablation, Adipocytes, Obese mouse model, Expression vectors, HSV-thymidine kinase
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