Advertisement

Quorum Sensing pp 235-249 | Cite as

Design of Synthetic Mammalian Quorum-Sensing Systems

  • Wilfried Weber
  • Martin FusseneggerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 692)

Abstract

Synthetic quorum-sensing systems in mammalian cells has enabled the implementation of time- and distance-dependent bioprocesses, as well as the design of synthetic ecosystems emulating clinically important host–parasite interactions. In this chapter, we provide a detailed protocol of the design of a mammalian cell-to-cell signaling device and its integration into a mammalian quorum-sensing system for cell density-induced expression product genes. Cell-to-cell signaling is based on a sender cell, metabolically engineered for expression of alcohol dehydrogenase converting ethanol into acetaldehyde, and a receiver cell line for the dose-dependent translation of the acetaldehyde concentration into transgene expression by an acetaldehyde-responsive promoter. This protocol can be adapted easily to various cell types and transgenes for the design of versatile mammalian cell-based quorum-sensing systems.

Key words

Acetaldehyde AIR Alcohol dehydrogenase Cell phone Synthetic biology Synthetic ecosystem 

Notes

Acknowledgment

We thank Marcia Schoenberg for critical comments on the manuscript. This work was supported by the Swiss National Science Foundation (grant no. 3100A0-112549).

References

  1. 1.
    You, L., Cox, R. S., III, Weiss, R., and Arnold, F. H. (2004) Programmed population control by cell–cell communication and regulated killing. Nature 428, 868–71.PubMedCrossRefGoogle Scholar
  2. 2.
    Basu, S., Gerchman, Y., Collins, C. H., Arnold, F. H., and Weiss, R. (2005) A synthetic multicellular system for programmed pattern formation. Nature 434, 1130–4.PubMedCrossRefGoogle Scholar
  3. 3.
    Balagadde, F. K., Song, H., Ozaki, J., Collins, C. H., Barnet, M., Arnold, F. H., Quake, S. R., and You, L. (2008) A synthetic Escherichia coli predator-prey ecosystem. Mol Syst Biol 4, 187.PubMedCrossRefGoogle Scholar
  4. 4.
    Weber, W., Daoud-El Baba, M., and Fussenegger, M. (2007) Synthetic ecosystems based on airborne inter- and intrakingdom communication. Proc Natl Acad Sci U S A 104, 10435–40.PubMedCrossRefGoogle Scholar
  5. 5.
    Weber, W., Rimann, M., Spielmann, M., Keller, B., Daoud-El Baba, M., Aubel, D., Weber, C. C., and Fussenegger, M. (2004) Gas-inducible transgene expression in mammalian cells and mice. Nat Biotechnol 22, 1440–4.PubMedCrossRefGoogle Scholar
  6. 6.
    Clemens, D. L., Forman, A., Jerrells, T. R., Sorrell, M. F., and Tuma, D. J. (2002) Relationship between acetaldehyde levels and cell survival in ethanol-metabolizing hepatoma cells. Hepatology 35, 1196–204.PubMedCrossRefGoogle Scholar
  7. 7.
    Doyle, A., Griffiths, J. B., and Newell, D. G. (1995) Cell & Tissue Culture: Laboratory Procedures, Vol. Update 9, Wiley, Hoboken, NJ.Google Scholar
  8. 8.
    Schlatter, S., Rimann, M., Kelm, J., and Fussenegger, M. (2002) SAMY, a novel mammalian reporter gene derived from Bacillus stearothermophilus alpha-amylase. Gene 282, 19–31.PubMedCrossRefGoogle Scholar
  9. 9.
    Fluri, D. A., Kelm, J. M., Lesage, G., Baba, M. D., and Fussenegger, M. (2007) InXy and SeXy, compact heterologous reporter proteins for mammalian cells. Biotechnol Bioeng 98, 655–67.PubMedCrossRefGoogle Scholar
  10. 10.
    Fluri, D. A., Kemmer, C., Daoud-El Baba, M., and Fussenegger, M. (2008) A novel system for trigger-controlled drug release from polymer capsules. J Control Release 131, 211–9.PubMedCrossRefGoogle Scholar
  11. 11.
    Hartenbach, S., and Fussenegger, M. (2005) Autoregulated, bidirectional and multicistronic gas-inducible mammalian as well as lentiviral expression vectors. J Biotechnol 120, 83–98.PubMedCrossRefGoogle Scholar
  12. 12.
    Weber, W., Rimann, M., de Glutz, F. N., Weber, E., Memmert, K., and Fussenegger, M. (2005) Gas-inducible product gene expression in bioreactors. Metab Eng 7, 174–81.PubMedCrossRefGoogle Scholar
  13. 13.
    Derouazi, M., Martinet, D., Besuchet Schmutz, N., Flaction, R., Wicht, M., Bertschinger, M., Hacker, D. L., Beckmann, J. S., and Wurm, F. M. (2006) Genetic characterization of CHO production host DG44 and derivative recombinant cell lines. Biochem Biophys Res Commun 340, 1069–77.PubMedCrossRefGoogle Scholar
  14. 14.
    Weber, W., and Fussenegger, M. (2004) Inducible gene expression in mammalian cells and mice. Methods Mol Biol 267, 451–66.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Biosystems Science and EngineeringETH ZurichBaselSwitzerland

Personalised recommendations