9. Native Promoter Reporters Validate Transcriptional Targets
| Abstract |
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The transcriptional activator β-catenin is the key mediator of the canonical Wnt signaling pathway. However, β-catenin does
not itself bind DNA, but functions via interaction with T-cell factor (TCF)/ lymphoid-enhancing factor (LEF) transcription
factors. These proteins contain a high-mobility group (HMG) box that binds DNA in a sequence-specific manner. Thus, in the
case of active Wnt signaling, β-catenin activates, in cooperation with proteins of the TCF/LEF family, the expression of a
wide variety of genes. To date, the list of established Wnt targets is far from complete. The establishment of plasmids harbouring
reporter genes under control of the native promoter sequences provides a tool to validate novel putative Wnt targets by directly
quantifying the β-catenin-dependent activation of each specific gene. In this chapter, we describe how to generate such reporter
plasmids using the MMP7 promoter as an example.
Series: Methods in Molecular Biology | Volume: 468 | Pub. Date: Jul-01-2008 | Page Range: 111-128 | DOI: 10.1007/978-1-59745-249-6_9
Subject: Protein Science
Key Words: Wnt pathway - Native promoter regions - TCF recognition sites - Reporter plasmids - Bacmid - Firefly luciferase
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