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The transience of transient overexpression

Nature Methods volume 10, pages 715721 (2013) | Download Citation

Much of what is known about mammalian cell regulation has been achieved with the aid of transiently transfected cells. However, overexpression can violate balanced gene dosage, affecting protein folding, complex assembly and downstream regulation. To avoid these problems, genome engineering technologies now enable the generation of stable cell lines expressing modified proteins at (almost) native levels.

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Acknowledgements

We thank many colleagues at the European Molecular Biology Laboratory, Monod Institute and two research consortia, the German National Genome Research Network–funded DiGtoP and European Union–funded SyBoSS—which are focused on developing and applying genome engineered cell lines—for useful discussions. We apologize for not citing many important references because of space limitations. M.S. is funded by DiGtoP. R.A.V. is supported by Centre National de la Recherche Scientifique, La Ligue contre le Cancer (Comité de Paris), l'Université Paris Diderot–Paris 7 and Institut Universitaire de France. Special thanks to I. Poser, M. Augsburg and A. Nitzsche (Max Planck Institute of Molecular Cell Biology and Genetics, Dresden) for providing the images of stably engineered cell lines.

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Affiliations

  1. Toby J. Gibson and Markus Seiler are in the Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

    • Toby J Gibson
    •  & Markus Seiler
  2. Reiner A. Veitia is at the Université Paris-Diderot–Paris 7 and Institut Universitaire de France and in Unité Mixte de Recherche 7592, Centre National de la Recherche Scientifique and Institut Jacques Monod, Paris, France.

    • Reiner A Veitia

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The authors declare no competing financial interests.

Corresponding authors

Correspondence to Toby J Gibson or Reiner A Veitia.

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https://doi.org/10.1038/nmeth.2534

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