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Mammalian cytochrome P-450s in the CYP1A gene family catalyse the oxidation of a wide range of drugs and foreign compounds resulting in their excretion. These enzymes are highly inducible by a range of compounds, including polycyclic aromatic hydrocarbons such as 3-methylcholanthrene (3-MC) and dioxins. Analysis of the CYP1A1 promoter has identified dioxin responsive enhancer elements which mediate the induction response. In order to evaluate this promoter as an in vivo regulatable expression system and to gain further insights into the tissue specific regulation of this gene, an 8.5 kb genomic fragment of the rat CYP1A1 promoter was cloned upstream of the lacZ reporter gene. This construct was used to generate transgenic mice and three independent lines were expanded for further study. The regulation of beta-galactosidase expression was determined in mock and 3-MC-treated mice in an extensive range of tissues. In untreated animals no transgene expression was detectable over non-transgenic controls. Treatment with 3-MC caused a profound increase in transgene expression (&amp;gt; 1,000-fold) in many tissues including liver, adrenal, kidney and intestine. Inducible transgene expression was also detectable in many of the other tissues including the spleen, lung, pancreas and the reproductive organs. Although the absolute levels of induction varied, no significant differences in the pattern of transgene expression were observed between the three different transgenic mouse lines. In addition, the pattern of transgene expression correlated closely with the reported regulation of CYP1A1 protein. These results indicate that the CYP1A1 promoter can drive expression of heterologous genes in a truly on/off manner in a variety of tissues and cell types which will allow the expression of other proteins to be controlled in vivo. This reporter system also provides a model for establishing the environmental and hormonal factors regulating the expression of the CYP1A1 gene.

Regulation of the CYP1A1 promoter in transgenic mice: an exquisitely sensitive on-off system for cell specific gene regulation