Abstract
In multicellular systems cell identity is imprinted by epigenetic regulation circuits, which determine the global transcriptome of adult cells in a cell phenotype–specific manner1,2,3. By combining two repressors, which control each other's expression, we have developed a mammalian epigenetic circuitry able to switch between two stable transgene expression states after transient administration of two alternate drugs. Engineered Chinese hamster ovary cells (CHO-K1) showed toggle switch–specific expression profiles of a human glycoprotein in culture, as well as after microencapsulation and implantation into mice. Switch dynamics and expression stability could be predicted with mathematical models. Epigenetic transgene control through toggle switches is an important tool for engineering artificial gene networks in mammalian cells.
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References
Rank, G., Prestel, M. & Paro, R. Transcription through intergenic chromosomal memory elements of the Drosophila bithorax complex correlates with an epigenetic switch. Mol. Cell. Biol. 22, 8026–8034 (2002).
Casadesus, J. & D'Ari, R. Memory in bacteria and phage. Bioessays 24, 512–518 (2002).
Kohler, C. & Grossniklaus, U. Epigenetic inheritance of expression states in plant development: the role of Polycomb group proteins. Curr. Opin. Cell Biol. 14, 773–779 (2002).
Malleret, G. et al. Inducible and reversible enhancement of learning, memory, and long-term potentiation by genetic inhibition of calcineurin. Cell 104, 675–686 (2001).
Aubel, D. et al. Design of a novel mammalian screening system for the detection of bioavailable, non-cytotoxic streptogramin antibiotics. J. Antibiot. 54, 44–55 (2001).
Fussenegger, M., Schlatter, S., Datwyler, D., Mazur, X. & Bailey, J.E. Controlled proliferation by multigene metabolic engineering enhances the productivity of Chinese hamster ovary cells. Nat. Biotechnol. 16, 468–472 (1998).
Rivera, V.M. et al. Long-term regulated expression of growth hormone in mice after intramuscular gene transfer. Proc. Natl. Acad. Sci. USA 96, 8657–8662 (1999).
Niwa, H., Miyazaki, J. & Smith, A.G. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nature Genet. 24, 372–376 (2000).
Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA 89, 5547–5551 (1992).
Yao, F. et al. Tetracycline repressor, tetR, rather than the tetR-mammalian cell transcription factor fusion derivatives, regulates inducible gene expression in mammalian cells. Hum. Gene Ther. 9, 1939–1950 (1998).
Fussenegger, M. et al. Streptogramin-based gene regulation systems for mammalian cells. Nat. Biotechnol. 18, 1203–1208 (2000).
Weber, W. et al. Macrolide-based transgene control in mammalian cells and mice. Nat. Biotechnol. 20, 901–907 (2002).
Weber, W. et al. Streptomyces-derived quorum-sensing systems engineered for adjustable transgene expression in mammalian cells and mice. Nucleic Acids Res. 31, e71 (2003).
Moosmann, P., Georgiev, O., Thiesen, H.J., Hagmann, M. & Schaffner, W. Silencing of RNA polymerases II and III-dependent transcription by the KRAB protein domain of KOX1, a Kruppel-type zinc finger factor. Biol. Chem. 378, 669–677 (1997).
Kapunisk-Uner, J.E., Sande, M.A. & Chambers, H.F.S. in Goodman and Gilman's The Pharmacological Basis of Therapeutics (ed. Limbierd, L.E.) 1123–1153 (McGraw-Hill, New York, 1996).
Wegener, H.C., Bager, F. & Aarestrup, F.M. Surveillance of antimicrobial resistance in humans, food stuffs and livestock in Denmark. Euro. Surveill. 2, 17–19 (1997).
Orlando, V. Polycomb, epigenomes, and control of cell identity. Cell 112, 599–606 (2003).
Hasty, J., McMillen, D. & Collins, J.J. Engineered gene circuits. Nature 420, 224–230 (2002).
Elowitz, M.B. & Leibler, S. A synthetic oscillatory network of transcriptional regulators. Nature 403, 335–338 (2000).
Atkinson, M.R., Savageau, M.A., Myers, J.T. & Ninfa, A.J. Development of genetic circuitry exhibiting toggle switch or oscillatory behavior in Escherichia coli. Cell 113, 597–607 (2003).
Gardner, T.S., Cantor, C.R. & Collins, J.J. Construction of a genetic toggle switch in Escherichia coli. Nature 403, 339–342 (2000).
Moser, S. et al. An update of pTRIDENT multicistronic expression vectors: pTRIDENTs containing novel streptogramin-responsive promoters. Biotechnol. Prog. 16, 724–735 (2000).
Weber, W. et al. Versatile macrolide-responsive mammalian expression vectors for multiregulated multigene metabolic engineering. Biotechnol. Bioeng. 80, 691–705 (2002).
Schlatter, S., Rimann, M., Kelm, J. & Fussenegger, M. SAMY, a novel mammalian reporter gene derived from Bacillus stearothermophilus alpha-amylase. Gene 282, 19–31 (2002).
Berger, J., Hauber, J., Hauber, R., Geiger, R. & Cullen, B.R. Secreted placental alkaline phosphatase: a powerful new quantitative indicator of gene expression in eukaryotic cells. Gene 66, 1–10 (1988).
Cherry, J.L. & Adler, F.R. How to make a biological switch. J. Theor. Biol. 203, 117–133 (2000).
Ermentrout, B. Simulating, Analyzing, and Animating Dynamical Systems (Society for Industrial & Applied Mathemetics, Pittsburgh, PA, 2002).
Corish, P. & Tyler-Smith, C. Attenuation of green fluorescent protein half-life in mammalian cells. Protein Eng. 12, 1035–1040 (1999).
Acknowledgements
We thank Martine Gilet for skillful assistance on in vivo experiments, Eva Niederer for FACS-mediated single-cell sorting, Valeria Gonzalez-Nicolini for useful discussions and Cornelia Fux for technical advice. This work was supported by the Swiss National Science Foundation (grant no. 631-065946) as well as the Novartis Foundation.
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Kramer, B., Viretta, A., Baba, ME. et al. An engineered epigenetic transgene switch in mammalian cells. Nat Biotechnol 22, 867–870 (2004). https://doi.org/10.1038/nbt980
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DOI: https://doi.org/10.1038/nbt980
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