( sily inexpensively andquantitativel...)Signal Transduction Reporting Systems Using Cis-Acting Enhancer,,,Elements

sily, inexpensively and quantitatively. Reporter enzyme assays replace tedious blotting procedures and are widely used due to their convenience, sensitivity and ability to generate reliable in vivo data.³

PathDetect Cis-Reporting Systems

figure 1

Each of the PathDetect in vivo signal transduction pathway cis-reporting systems includes one of five cis- reporter plasmids and a positive control plasmid. Positive control plasmids express gene products known to activate the signaling pathway converging at the corresponding enhancer elements. The five reporter plasmids (figure 1) of the PathDetect cis-reporting systems contain tandem repeats of one of the following enhancer elements: cyclic AMP response element (CRE), serum response element (SRE), nuclear factor kB (NF-kB), activator protein 1 (AP-1) or serum response factor (SRF). Together with a TATA box, these enhancer elements control the expression of the luciferase reporter gene located downstream.

Since most enhancer elements can be regulated by more than one transcription activator, each enhancer element can be used to monitor more than one signal transduction pathway. When a plasmid expressing a gene of interest is cotransfected into mammalian cells along with one of these cis-reporter plasmids, increased luciferase activity indicates transcription activation. The PathDetect cis-reporting systems are useful tools for assessing the potential involvement of a gene product in a pathway. For more specific assessment, the PathDetect trans-reporting systems can be used.⁴

Response to Overexpression of Known Activators

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