Inducible, tightly regulated and non-leaky neuronal gene expression in mice
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.
KeywordsTET system Transgenic mice Doxycycline LacZ reporter Leakage Thy1.2 promoter
- Dewachter I, Reverse D, Caluwaerts N, Ris L, Kuiperi C, Van den Haute C, Spittaels K, Umans L, Serneels L, Thiry E, Moechars D, Mercken M, Godaux E, Van Leuven F (2002) Neuronal deficiency of presenilin 1 inhibits amyloid plaque formation and corrects hippocampal long-term potentiation but not a cognitive defect of amyloid precursor protein [V717I] transgenic mice. J Neurosci Off J Soc Neurosci 22(9):3445–3453. doi:20026290 Google Scholar
- Duerr J, Gruner M, Schubert SC, Haberkorn U, Bujard H, Mall MA (2011) Use of a new-generation reverse tetracycline transactivator system for quantitative control of conditional gene expression in the murine lung. Am J Respir Cell Mol Biol 44(2):244–254. doi:10.1165/rcmb.2009-0115OC PubMedCrossRefGoogle Scholar
- Jones J, Nivitchanyong T, Giblin C, Ciccarone V, Judd D, Gorfien S, Krag SS, Betenbaugh MJ (2005) Optimization of tetracycline-responsive recombinant protein production and effect on cell growth and ER stress in mammalian cells. Biotechnol Bioeng 91(6):722–732. doi:10.1002/bit.20566 PubMedCrossRefGoogle Scholar
- Urlinger S, Baron U, Thellmann M, Hasan MT, Bujard H, Hillen W (2000) Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc Natl Acad Sci USA 97(14):7963–7968. doi:10.1073/pnas.130192197 PubMedCentralPubMedCrossRefGoogle Scholar