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An engineered epigenetic transgene switch in mammalian cells

Nature Biotechnology volume 22, pages 867–870 (2004)Cite this article

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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Figure 1: Molecular configuration of the epigenetic transcription control circuit.
Figure 2: Validation of bistable toggle expression performance after expression in CHO-K1 cells.
Figure 3: Reversibility of epigenetic expression control in TOGGLECHO37.
Figure 4: Bistability of epigenetic expression control in mice.

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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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  1. Alessandro Usseglio Viretta

    Present address: Institute of Robotics and Intelligent Systems, ETH Zurich, CH-8092, Zurich, Switzerland

Authors and Affiliations

  1. Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich, Zurich, CH-8093, Switzerland

    Beat P Kramer, Alessandro Usseglio Viretta, Wilfried Weber & Martin Fussenegger

  2. Département Génie Biologique, Institut Universitaire de Technologie, IUTA, Villeurbanne, F-69622, Cedex, France

    Marie Daoud-El Baba & Dominique Aubel

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  1. Beat P Kramer
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Correspondence to Martin Fussenegger.

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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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