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.
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.
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