Abstract
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.
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Acknowledgments
This work was supported by grants to TB from the German Research Ministry BMBF (NGFN2 project no. 01GS0436). SS was funded by the German Research Council DFG (grant no. BR1399/4-3) and OS was funded by the Deutsche Krebshilfe (grant no. 106958).
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Schmalhofer, O., Spaderna, S., Brabletz, T. (2008). Native Promoter Reporters Validate Transcriptional Targets. In: Vincan, E. (eds) Wnt Signaling. Methods in Molecular Biology™, vol 468. Humana Press. https://doi.org/10.1007/978-1-59745-249-6_9
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DOI: https://doi.org/10.1007/978-1-59745-249-6_9
Publisher Name: Humana Press
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