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Functional cross-kingdom conservation of mammalian and moss (Physcomitrella patens) transcription, translation and secretion machineries

Authors

  • Marc Gitzinger,

    1. Department for Biosystems Science and Engineering, ETH Zurich, Wolfgang-Pauli-Strasse 10, HCI F115, CH-8093 Zurich, Switzerland
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  • Juliana Parsons,

    1. Plant Biotechnology, Faculty of Biology, and Centre for Biological Signalling Studies (BIOSS), University of Freiburg, Schaenzlestr. 1, D-79104 Freiburg, Germany
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  • Ralf Reski,

    1. Plant Biotechnology, Faculty of Biology, and Centre for Biological Signalling Studies (BIOSS), University of Freiburg, Schaenzlestr. 1, D-79104 Freiburg, Germany
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  • Martin Fussenegger

    Corresponding author
    1. Department for Biosystems Science and Engineering, ETH Zurich, Wolfgang-Pauli-Strasse 10, HCI F115, CH-8093 Zurich, Switzerland
      * Correspondence (fax +41 44 633 12 34; e-mail: fussenegger@chem.ethz.ch)
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Errata

This article is corrected by:

  1. Errata: Corrigendum Volume 7, Issue 7, 717, Article first published online: 11 August 2009
  2. Errata: Corrigendum Volume 7, Issue 2, 210, Article first published online: 8 January 2009

* Correspondence (fax +41 44 633 12 34; e-mail: fussenegger@chem.ethz.ch)

Summary

Plants and mammals are separated by a huge evolutionary distance. Consequently, biotechnology and genetics have traditionally been divided into ‘green’ and ‘red’. Here, we provide comprehensive evidence that key components of the mammalian transcription, translation and secretion machineries are functional in the model plant Physcomitrella patens. Cross-kingdom compatibility of different expression modalities originally designed for mammalian cells, such as native and synthetic promoters and polyadenylation sites, viral and cellular internal ribosome entry sites, secretion signal peptides and secreted product proteins, and synthetic transactivators and transrepressors, was established. This mammalian expression portfolio enabled constitutive, conditional and autoregulated expression of different product genes in a multicistronic expression format, optionally adjusted by various trigger molecules, such as butyrolactones, macrolide antibiotics and ethanol. Capitalizing on a cross-kingdom-compatible expression platform, we pioneered a prototype biopharmaceutical manufacturing scenario using microencapsulated transgenic P. patens protoplasts cultivated in a Wave Bioreactor. Vascular endothelial growth factor 121 (VEGF121) titres matched those typically achieved by standard protonema populations grown in stirred-tank bioreactors. The full compatibility of mammalian expression systems in P. patens further promotes the use of moss as a cost-effective alternative for the manufacture of complex biopharmaceuticals, and as a valuable host system to advance synthetic biology in plants.

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