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Tet-system for the regulation of gene expression during embryonic development

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Abstract

The ability to control gene expression in a temporal and spatial manner provides a new tool for the study of mammalian gene function particularly during development and oncogenesis. In this study the suitability of the tet-system for investigating embryogenesis was tested in detail. The tTA CMV (M1) and rTA CMV-3 (reverse Tc-controlled transactivator) transgenic mice were bred with NZL-2 bi-reporter mice containing the vector with a tTA/rTA responsive bidirectional promoter that allows simultaneous regulation of expression of two reporter genes encoding luciferase and β-galactosidase. In both cases reporter genes were found to be expressed in a wide spectrum of tissues of double transgenic embryos and adult mice. The earliest expression was detected in tTA CMV (M1)/NZL-2 embryos at embryonic day 10.5 (E10.5) and rTA CMV -3/NZL-2 embryos at E13.5. Doxycycline abolished β-gal expression in tTA CMV (M1)/NZL-2 but induced it in rTA CMV -3/NZL-2 embryos including late stages of embryogenesis. The tTA and rtTA transactivators thus revealed a partially complementary mode of action during second half of embryonic development. These experiments demonstrated that both Tet regulatory systems function during embryonic development. We conclude that the Tet systems allows regulation of gene expression during embryonic development and that ‘double reporter’ animals like the NZL-2 mice are useful tools for the characterization of newly generated tet transactivator lines expressing tTA (or rtTA) in embryonic as well as in adult tissues.

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Authors and Affiliations

  1. Institut für Medizinische Strahlenkunde und Zellforschung, Bayerische Julius-Maximilians-Universität Würzburg, Versbacher-Str. 5, D-97078, Würzburg, Germany

    Lev M. Fedorov, Oleg Yu. Tyrsin, Veit Krenn & Ulf R. Rapp

  2. Institut für Med. Strahlenkunde und Zellforschung (MSZ), Bayerische Julius- Maximilians-Universität, Versbacher Str. 5, D-97078, Würzburg, Germany

    Lev M. Fedorov & Ulf R. Rapp

  3. Sechenov Institute of Evolutionary Physiology and Biochemistry, RAN, 44 Thorez pr., 194223, Sankt-Petersburg, Russia

    Elena V. Chernigovskaya

Authors
  1. Lev M. Fedorov
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  2. Oleg Yu. Tyrsin
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  3. Veit Krenn
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  4. Elena V. Chernigovskaya
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  5. Ulf R. Rapp
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Fedorov, L.M., Tyrsin, O.Y., Krenn, V. et al. Tet-system for the regulation of gene expression during embryonic development. Transgenic Res 10, 247–258 (2001). https://doi.org/10.1023/A:1016632110931

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