Conditional and constitutive expression of a Tbx1-GFP fusion protein in mice

Laina Freyer, Sonja Nowotschin, Melinda K. Pirity, Antonio Baldini, Bernice E. Morrow

Research output: Contribution to journalArticle

Abstract

Background: Velo-cardio-facial syndrome/DiGeorge syndrome (VCFS/DGS) is caused by a 1.5-3 Mb microdeletion of chromosome 22q11.2, frequently referred to as 22q11.2 deletion syndrome (22q11DS). This region includes TBX1, a T-box transcription factor gene that contributes to the etiology of 22q11DS. The requirement for TBX1 in mammalian development is dosage-sensitive, such that loss-of-function (LOF) and gain-of-function (GOF) of TBX1 in both mice and humans results in disease relevant congenital malformations. Results: To further gain insight into the role of Tbx1 in development, we have targeted the Rosa26 locus to generate a new GOF mouse model in which a Tbx1-GFP fusion protein is expressed conditionally using the Cre/LoxP system. Tbx1-GFP expression is driven by the endogenous Rosa26 promoter resulting in ectopic and persistent expression. Tbx1 is pivotal for proper ear and heart development; ectopic activation of Tbx1-GFP in the otic vesicle by Pax2-Cre and Foxg1-Cre represses neurogenesis and produces morphological defects of the inner ear. Overexpression of a single copy of Tbx1-GFP using Tbx1 §ssup§ Cre/+ §esup§ was viable, while overexpression of both copies resulted in neonatal lethality with cardiac outflow tract defects. We have partially rescued inner ear and heart anomalies in Tbx1 §ssup§ Cre/- §esup§ null embryos by expression of Tbx1-GFP. Conclusions: We have generated a new mouse model to conditionally overexpress a GFP-tagged Tbx1 protein in vivo. This provides a useful tool to investigate in vivo direct downstream targets and protein binding partners of Tbx1.

Original languageEnglish
Article number33
JournalBMC Developmental Biology
Volume13
Issue number1
DOIs
Publication statusPublished - 2013

Fingerprint

DiGeorge Syndrome
Inner Ear
Ear
Proteins
Neurogenesis
Protein Binding
Transcription Factors
Embryonic Structures
Chromosomes
Genes

Keywords

  • Gain-of-function
  • Mouse model
  • Rosa26
  • Tbx1
  • VCFS/DGS

ASJC Scopus subject areas

  • Developmental Biology
  • Medicine(all)

Cite this

Freyer, L., Nowotschin, S., Pirity, M. K., Baldini, A., & Morrow, B. E. (2013). Conditional and constitutive expression of a Tbx1-GFP fusion protein in mice. BMC Developmental Biology, 13(1), [33]. https://doi.org/10.1186/1471-213X-13-33

Conditional and constitutive expression of a Tbx1-GFP fusion protein in mice. / Freyer, Laina; Nowotschin, Sonja; Pirity, Melinda K.; Baldini, Antonio; Morrow, Bernice E.

In: BMC Developmental Biology, Vol. 13, No. 1, 33, 2013.

Research output: Contribution to journalArticle

Freyer, L, Nowotschin, S, Pirity, MK, Baldini, A & Morrow, BE 2013, 'Conditional and constitutive expression of a Tbx1-GFP fusion protein in mice', BMC Developmental Biology, vol. 13, no. 1, 33. https://doi.org/10.1186/1471-213X-13-33
Freyer, Laina ; Nowotschin, Sonja ; Pirity, Melinda K. ; Baldini, Antonio ; Morrow, Bernice E. / Conditional and constitutive expression of a Tbx1-GFP fusion protein in mice. In: BMC Developmental Biology. 2013 ; Vol. 13, No. 1.
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AB - Background: Velo-cardio-facial syndrome/DiGeorge syndrome (VCFS/DGS) is caused by a 1.5-3 Mb microdeletion of chromosome 22q11.2, frequently referred to as 22q11.2 deletion syndrome (22q11DS). This region includes TBX1, a T-box transcription factor gene that contributes to the etiology of 22q11DS. The requirement for TBX1 in mammalian development is dosage-sensitive, such that loss-of-function (LOF) and gain-of-function (GOF) of TBX1 in both mice and humans results in disease relevant congenital malformations. Results: To further gain insight into the role of Tbx1 in development, we have targeted the Rosa26 locus to generate a new GOF mouse model in which a Tbx1-GFP fusion protein is expressed conditionally using the Cre/LoxP system. Tbx1-GFP expression is driven by the endogenous Rosa26 promoter resulting in ectopic and persistent expression. Tbx1 is pivotal for proper ear and heart development; ectopic activation of Tbx1-GFP in the otic vesicle by Pax2-Cre and Foxg1-Cre represses neurogenesis and produces morphological defects of the inner ear. Overexpression of a single copy of Tbx1-GFP using Tbx1 §ssup§ Cre/+ §esup§ was viable, while overexpression of both copies resulted in neonatal lethality with cardiac outflow tract defects. We have partially rescued inner ear and heart anomalies in Tbx1 §ssup§ Cre/- §esup§ null embryos by expression of Tbx1-GFP. Conclusions: We have generated a new mouse model to conditionally overexpress a GFP-tagged Tbx1 protein in vivo. This provides a useful tool to investigate in vivo direct downstream targets and protein binding partners of Tbx1.

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