Assaying β-Catenin/TCF Transcription with β-Catenin/TCF Transcription-Based Reporter Constructs (original) (raw)

Abstract

Transcription-based reporters have been instrumental in characterizing the Wnt/β-catenin signaling pathway and will be essential in the search for therapeutics aimed at combating diseases linked to aberrant signaling. In this chapter, we introduce a new improved Wnt/β-catenin reporter system, β-catenin-activated reporter (BAR), and its accompanying control reporter system, found unresponsive BAR (fuBAR). Its enhanced sensitivity, increased dynamic range, and lentiviral platform provide a reporter system that will keep pace with the needs of scientists in the field.

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References

  1. Nusslein-Volhard, C., and Wieschaus, E. (1980). Mutations affecting segment number and polarity in Drosophila. Nature 287, 795–801.
    Article Google Scholar
  2. McMahon, A.P. and Moon, R.T. (1989). Ectopic expression of the proto-oncogene int-1 in Xenopus embryos leads to duplication of the embryonic axis. Cell 58, 1075–1084.
    Article CAS PubMed Google Scholar
  3. Jue, S.F., Bradley, R.S., Rudnicki, J.A., Varmus, H.E., and Brown, A.M. (1992). The mouse Wnt-1 gene can act via a paracrine mechanism in transformation of mammary epithelial cells. Mol Cell Biol 12, 321–328.
    CAS PubMed Google Scholar
  4. Korinek, V., Barker, N., Morin, P.J., van Wichen, D., de Weger, R., Kinzler, K.W., Vogelstein, B., and Clevers, H. (1997). Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC -/- colon carcinoma. Science 275, 1784 –1787.
    Article CAS PubMed Google Scholar
  5. Veeman, M.T., Slusarski, D.C., Kaykas, A., Louie, S.H., and Moon, R.T. (2003). Zebrafish prickle, a modulator of non-canonical Wnt/Fz signaling, regulates gastrulation movements. Curr Biol 13, 680–685.
    Article CAS PubMed Google Scholar
  6. DasGupta, R., Kaykas, A., Moon, R.T., and Perrimon, N. (2005). Functional genomic analysis of the Wnt-wingless signaling pathway. Science 308, 826–833.
    Article CAS PubMed Google Scholar
  7. Rekas, A., Alattia, J.R., Nagai, T., Miyawaki, A., and Ikura, M. (2002). Crystal structure of Venus, a yellow fluorescent protein with improved maturation and reduced environmental sensitivity. J Biol Chem 277, 50573–50578.
    Article CAS PubMed Google Scholar
  8. Barolo, S., and Posakony, J.W. (2002). Three habits of highly effective signaling pathways: principles of transcriptional control by developmental cell signaling. Genes Dev 16, 1167–1181.
    Article CAS PubMed Google Scholar
  9. Olson, L.E., Tollkuhn, J., Scafoglio, C., Krones, A., Zhang, J., Ohgi, K.A., Wu, W., Taketo, M.M., Kemler, R., Grosschedl, R., et al. (2006). Homeodomain-mediated beta-catenin-dependent switching events dictate cell-lineage determination. Cell 125, 593–605.
    Article CAS PubMed Google Scholar
  10. Willert, K., Brown, J. D., Danenberg, E., Duncan, A. W., Weissman, I. L., Reya, T., et al. (2003) Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature, 423, 448–452.
    Article CAS PubMed Google Scholar

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Acknowledgments

We thank the Howard Hughes Medical Institute for funding.

Author information

Authors and Affiliations

  1. Howard Hughes Medical Institute and Department of Pharmacology and Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, WA, USA
    Travis L. Biechele & Randall T. Moon

Authors

  1. Travis L. Biechele
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  2. Randall T. Moon
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Editor information

Editors and Affiliations

  1. University of Melbourne, Parkville, Victoria, VIC 3010, Australia
    Elizabeth Vincan PhD

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Biechele, T.L., Moon, R.T. (2008). Assaying β-Catenin/TCF Transcription with β-Catenin/TCF Transcription-Based Reporter Constructs. In: Vincan, E. (eds) Wnt Signaling. Methods in Molecular Biology™, vol 468. Humana Press. https://doi.org/10.1007/978-1-59745-249-6\_8

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