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Related Experiment Video

Updated: Dec 10, 2025

In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila
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Functionally distinct roles for T and Tbx6 during mouse development.

Amy K Wehn1, Deborah R Farkas1, Carly E Sedlock1

  • 1Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.

Biology Open
|August 29, 2020
PubMed
Summary
This summary is machine-generated.

Mouse T-box transcription factors T and Tbx6 have distinct DNA binding affinities and transcriptional activities. These differences explain why Tbx6 cannot functionally replace T and may lead to competition in vivo.

Keywords:
BrachyuryMesodermMouseTT-boxTbx6

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Area of Science:

  • Developmental Biology
  • Molecular Genetics
  • Gene Regulation

Background:

  • T-box transcription factors T and Tbx6 are co-expressed in mouse primitive streak.
  • T is crucial for notochord development, while Tbx6 is essential for presomitic mesoderm development.
  • Both factors share a DNA-binding T-domain, suggesting potential functional overlap.

Purpose of the Study:

  • To investigate the functional similarities and differences in DNA binding and transcriptional activity of T and Tbx6.
  • To analyze the genetic interactions and competition between T and Tbx6 in vivo.
  • To determine if Tbx6 can functionally substitute for T.

Main Methods:

  • In vitro DNA binding and transcriptional assays using T-domains of T and Tbx6.
  • Generation of mouse embryos with altered Tbx6 expression levels (overexpression, heterozygous).
  • Genetic analysis using dominant-negative TWis allele and Tbx6 hypomorphic allele (rib-vertebrae).
  • Tbx6 knockin at the T locus in embryonic stem cells for chimeric embryo analysis.

Main Results:

  • T and Tbx6 exhibit different affinities for DNA binding sites at the Dll1 enhancer.
  • Tbx6 shows tenfold higher transcriptional activation than T in vitro.
  • Competition between T and Tbx6 for target gene enhancers requires functional DNA binding domains.
  • Tbx6 overexpression leads to short tails, mimicking T heterozygous phenotype.
  • Tbx6+/- TWis/+ embryos display rib fusions and vertebral malformations, similar to Tbx6 hypomorphic phenotypes.
  • Tbx6 knockin at the T locus results in T null-like phenotypes in chimeric embryos.

Conclusions:

  • Differences in DNA binding affinity and transcriptional activity between T and Tbx6 provide a molecular basis for their distinct roles.
  • Tbx6 cannot functionally replace T due to these molecular differences.
  • Competition between T and Tbx6 in vivo may contribute to observed developmental phenotypes.