Transgenic Research

, Volume 23, Issue 2, pp 225–233 | Cite as

Inducible, tightly regulated and non-leaky neuronal gene expression in mice

Original Paper

Abstract

The Tetracycline (Tet)-controlled inducible system is the most widely used reversible system for transgene expression in mice with over 500 lines created to date. Although this system has been optimized over the years, it still has limitations such as residual transgene expression when turned off, referred to as leakiness. Here, we present a series of new Tet-OFF transgenic mice based on the second generation tetracycline-responsive transactivator system. The tTA-Advanced (tTA2S) is expressed under control of the neuron-specific Thy1.2 promoter (Thy-OFF), to regulate expression in the mouse brain. In addition, we generated a lacZ reporter line, utilizing the Ptight Tet-responsive promoter (Ptight–lacZ), to test our system. Two Thy-OFF transgenic lines displaying two distinct patterns of expression were selected. Oral doxycycline treatment of Thy-OFF/Ptight–lacZ mice demonstrated tight transgene regulation with no leak expression. These new Thy-OFF mice are valuable for studies in a broad range of neurodegenerative diseases such as Alzheimer’s disease and related forms of dementia, where control of transgene expression is critical to understanding mechanisms underlying the disease. Furthermore, Ptight–lacZ reporter mice may be widely applicable.

Keywords

TET system Transgenic mice Doxycycline LacZ reporter Leakage Thy1.2 promoter 

Supplementary material

11248_2013_9767_MOESM1_ESM.pdf (509 kb)
Supplementary Fig. 1 Expression patterns of the four different reporter lines generated. X-Gal staining of the brain of the four initial different reporter lines generated revealed most widespread staining for line #31. All lines have been crossed with the same inducer line (Thy-OFF15) for comparison (PDF 508 kb)
11248_2013_9767_MOESM2_ESM.pdf (862 kb)
Supplementary Fig. 2 X-Gal staining of various organs reveals the specificity of the thy1.2 promoter. X-Gal staining of spinal cord, heart, liver and muscle of the wiltype (A), Ptight–lacZ31 (B), Thy-OFF6 (C) and Thy-OFF 15 (C), did not reveal expression. In contrast, Thy-OFF6/Ptight–lacZ31 and Thy-OFF15/Ptight–lacZ31 mice exhibit strong β-galactosidas expression in the spinal cord only. Interestingly, Thy-cre/R26R-lacZ mice show some expression outside the CNS (PDF 862 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Fabien Delerue
    • 1
    • 2
    • 3
  • Michael White
    • 4
  • Lars M. Ittner
    • 1
    • 2
    • 3
    • 5
  1. 1.Transgenic Animal Unit, School of Medical ScienceUniversity of New South WalesSydneyAustralia
  2. 2.Dementia Research Unit, School of Medical ScienceUniversity of New South WalesSydneyAustralia
  3. 3.Alzheimer’s and Parkinson’s Disease Laboratory, Brain and Mind Research InstituteUniversity of SydneySydneyAustralia
  4. 4.Department of PathologyWashington University School of MedicineSt LouisUSA
  5. 5.Neuroscience Research AustraliaSydneyAustralia

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