Cholesterol 7.alpha.-hydroxylase gene regulatory elements and methods for using them
AssigneeNortheastern Ohio Universities
InventorJohn Y. L. Chiang
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Abstract
DNA regulatory elements that control cholesterol 7.alpha.-hydroxylase expression are disclosed. A gene construct is provided comprising at least one regulatory element and a reporter gene is used in an assay to detect a compound that modulates cholesterol 7.alpha.-hydroxylase enzyme regulation. Thus, a method for screening compounds that inhibit or stimulate expression of the enzyme is provided, as well as a method for detecting and isolating the transcription factors of the cholesterol 7.alpha.-hydroxylase gene.
Claims16
An isolated regulatory element of the cholesterol 7.alpha.-hydroxylase (CYP7) gene which consists of a DNA fragment selected from the group consisting of DNA fragments from about -191 to about +64 of the rat CYP7 gene, from about -252 to about +3 of the hamster CYP7 gene and from about -187 to about +65 of the human CYP7 gene, wherein said DNA fragment is regulatory with respect to CYP7 expression in the presence of bile acids, and wherein the numbering of said DNA fragment is measured relative to the transcription start site.
An isolated regulatory element of the rat CYP7 gene which consists of a DNA fragment selected from the group consisting of DNA fragments from about -101 to about -29, from about -81 to about -37, wherein said DNA fragment is regulatory with respect to CYP7 expression in the presence of bile acids, and wherein the numbering of said DNA fragment is measured relative to the transcription start site.
An isolated regulatory element of the human CYP7 gene which consists of a DNA fragment selected from the group consisting of DNA fragments from about -104 to about -30, and from about -78 to about -36, wherein said DNA fragment is regulatory with respect to CYP7 expression in the presence of bile acids, and wherein the numbering of said DNA fragment is measured relative to the transcription start site.
An isolated regulatory element of the hamster CYP7 gene which consists of a DNA fragment selected from the group consisting of DNA fragments from about -161 to about -86, and from about -136 to about -92, wherein said DNA fragment is regulatory with respect to CYP7 expression in the presence of bile acids, and wherein the numbering of said DNA fragment is measured relative to the transcription start site.
A method for determining whether an agent inhibits or stimulates CYP7 gene expression comprising the steps of:
- (a) contacting said agent with a transfected cell containing a construct, wherein said construct comprises
- (i) human cholesterol 7.alpha.-hydroxylase (CYP7) DNA that consists of an isolated regulatory element located within a region from -187 to +65, wherein said element comprises nucleotides -78 to -36 and said element is regulatory with respect to CYP7 expression in the presence of bile acids, and wherein the numbering of said element is measured relative to the transcription start site and
- (ii) heterologous DNA, and
- (b) determining CYP7 gene expression in said transfected cell.
Dependencies are inferred from the claim text.