Tight regulation of transgene expression by tetracycline-dependent activator and repressor in brain

S. Uchida, S. Sakai, T. Furuichi, H. Hosoda, K. Toyota, T. Ishii, A. Kitamoto, M. Sekine, K. Koike, S. Masushige, G. Murphy, A. J. Silva, S. Kida

研究成果: Article査読

23 被引用数 (Scopus)

抄録

Methods to temporally and spatially regulate gene mutations will provide a powerful strategy to investigate gene function in the brain. To develop these methods, we have established a tightly regulated system for transgene expression in the forebrain using both a tetracycline (Tc)-dependent transcription activator (rtTA) and a repressor (TetR-Kruppel-associated box). In this system, the repressor binds to the Tc-responsive element (TRE) in the absence of doxycycline (Dox), leading to the repression of leaky activation of TRE-mediated transcription caused by weak binding of rtTA to TRE. Upon Dox administration, only the activator binds to TRE and activates transcription. We tested this system in cultured cells by bicistronically expressing both the regulators using an internal ribosome entry site (IRES). In COS-1, HeLa and SHSY5Y cells, leaky transcription activation led by rtTA in the absence of Dox was repressed without decreasing the level of activated transcription in the presence of Dox. Using this system, transgenic mice were produced that express both the regulators using IRES in the forebrain under the control of the αCaMKII promoter and were bred with transgenic mice carrying the TRE-dependent reporter transgene. In reverse transcription-polymerase chain reaction and in situ hybridization analyses of the forebrain in adult double transgenic mice, the treatment of Dox induces reporter mRNA expression, which was not detected before the treatment and after the withdraw of Dox following the treatment. These results indicate that this system allows the tight regulation of transgene expression in a Dox-dependent fashion in the forebrain and will be useful in investigating gene function in the brain.

本文言語English
ページ(範囲)96-106
ページ数11
ジャーナルGenes, Brain and Behavior
5
1
DOI
出版ステータスPublished - 2006 2月
外部発表はい

ASJC Scopus subject areas

  • 遺伝学
  • 神経学
  • 行動神経科学

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