Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells

Leuchtenberger S, Perz A, Gatz C, Bartsch JW (2001)
Nucleic Acids Research 29(16): 76e-76.

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Abstract
Conditional expression of suicide genes in vivo has a wide range of applications in biological research and requires a minimal basal promoter activity in the uninduced state. To reduce basal activity of tetracycline (tc)-inducible target promoters we combined synthetic tet operators in varying numbers with a core promoter derived from the plant viral 35S promoter. An optimized promoter, P_TF, was found to exert a stringent regulation of luciferase in combination with tTA and rtTA in different mammalian cell lines. We linked P_TF to the barnase gene, coding for a highly active RNase from Bacillus amyloliquefaciens. Stable cell clones expressing barnase under control of tTA exerted cell death only after tc withdrawal, correlating with a 10-fold induction of barnase mRNA expression. Directing tTA expression through a neuron-specific enolase promoter (P_NSE) leads to barnase expression and cell death in neuronal cells after tc withdrawal. Taken together, our data demonstrate that a stringent control of barnase expression in the uninduced state improves cell ablation studies, as high frequencies of transgene propagation in both cell lines and in transgenic mice are observed.
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Leuchtenberger S, Perz A, Gatz C, Bartsch JW. Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells. Nucleic Acids Research. 2001;29(16):76e-76.
Leuchtenberger, S., Perz, A., Gatz, C., & Bartsch, J. W. (2001). Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells. Nucleic Acids Research, 29(16), 76e-76.
Leuchtenberger, S., Perz, A., Gatz, C., and Bartsch, J. W. (2001). Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells. Nucleic Acids Research 29, 76e-76.
Leuchtenberger, S., et al., 2001. Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells. Nucleic Acids Research, 29(16), p 76e-76.
S. Leuchtenberger, et al., “Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells”, Nucleic Acids Research, vol. 29, 2001, pp. 76e-76.
Leuchtenberger, S., Perz, A., Gatz, C., Bartsch, J.W.: Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells. Nucleic Acids Research. 29, 76e-76 (2001).
Leuchtenberger, Stefanie, Perz, Angela, Gatz, Christiane, and Bartsch, Jörg W. “Conditional cell ablation by stringent tetracycline-dependent regulation of barnase in mammalian cells”. Nucleic Acids Research 29.16 (2001): 76e-76.
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