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Plasmid 12456: M50 Super 8x TOPFlash
  • Beta-catenin reporter. TCF/LEF sites upstream of a luciferase reporter.

  • TCF/LEF binding sites

  • beta catenin reporter

  • 120

  • Ctnnb1 (Bfc, Catnb, Mesc)

  • pTA-Luc
    (Search Vector Database)

  • Clontech

  • Luciferase

  • 4871

  • MluI

  • Unknown

  • MluI

  • Unknown

  • RVPrimer3 List of Sequencing Primers

  • LucNRev

  • Ampicillin

  • DH5alpha

  • 37

  • High Copy

  • The idea for the construct is from Hans Clevers lab, who designed the original TOPflash.

  • View sequences (3)
  • View map

  • Randall Moon

  • UBMTA
    Luciferase Limited Use Label License

Comments: 

This is a luciferase reporter of beta-catenin-mediated transcriptional activation. In HEK cells, maximal activation of this reporter is ~100-fold (activation by Wnt) up to ~1,000-fold (activation by phosphorylation mutants of beta-catenin). The appropriate control plasmid is clone M51, Super8XFOPflash, which has mutant TCF/LEF binding sites.

This construct was made by Ajamete Kaykas in the Moon lab. The backbone is the pTA-Luc vector of Clontech, which provides a minimal TA viral promoter driving expression of the firefly luciferase gene (see company publications for details). 7 TCF/LEF binding sites were cloned into the Mlu1 site of this vector (7 copies of: AGATCAAAGGgggta, with TCF/LEF binding site in CAP letters, and a spacer in lower case, separating each copy of the TCF/LEF site).

Note: This plasmid was published as M50 Super 8x TOPFlash, but the plasmid actually contains 7 TCF/LEF sites.

Addgene has sequenced a portion of this plasmid for verification. Full plasmid sequence is available only if provided by the depositing laboratory.

Article: Zebrafish prickle, a modulator of noncanonical Wnt/Fz signaling, regulates gastrulation movements. Veeman et al (Curr Biol. 2003 Apr 15. 13(8):680-5. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 12456" in your Materials and Methods section.