Exploring Cellular Signaling with Luciferase Reporters

• Current understanding of consensus binding sequences for transcription and other factors that regulate gene expression allows the creation of "synthetic promoter" elements.
• Specific signaling pathways can be monitored using luciferase under the control of these synthetic promoters.
• Using reporters to indicate signaling pathway activation is useful in studies involving small molecule inhibitors of signaling or siRNA knockdown experiments.

Here's how it works:

• Place multiple repeats of the desired consensus response element upstream of a minimal promoter.
• Use this synthetic promoter to drive luciferase reporter expression when the signaling pathway is activated.

For example:
If you are studying a G∝^S-associated GPCR pathway, you can use a construct in which luciferase expression is under the control of a minimal promoter coupled to a cAMP response element (CRE) (e.g., pGL4.29). Cells will respond with luciferase expression whenever the cAMP pathway is activated.

Evaluating Control of the NF-κB Pathway in B-Cell Lymphoma

Many B-cell lymphomas have a mutation in the A20 gene, regulator of the NF-κB pathway. The authors of this study used an NF-κB pathway pGL4 Vector (pGL4.32) to investigate the ability of wild type A20 or cancer-associated A20 mutant to control the NF-κB pathway in a B-cell lymphoma cell line.

Kato, M. et al. (2009) Nature 459, 712–6.

Tools

• Signaling Pathways Tools 
• Signaling Pathway Analysis Applications 
• Dual Luminescent Reporter Assay Tools

Above downloads in PDF format