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Gene Targeting for Precision Glyco-Engineering: Production of Biopharmaceuticals Devoid of Plant-Typical Glycosylation in Moss Bioreactors

  • Eva L. Decker
  • Gertrud Wiedemann
  • Ralf Reski
Part of the Methods in Molecular Biology book series (MIMB, volume 1321)

Abstract

One of the main challenges for the production of biopharmaceuticals in plant-based systems is the modulation of plant-specific glycosylation patterns towards a humanized form. Posttranslational modifications in plants are similar to those in humans, but several differences affect product quality and efficacy and can also cause immune responses in patients. In the moss Physcomitrella patens highly efficient gene targeting via homologous recombination enables glyco-engineering to obtain suitable platform lines for the production of recombinant proteins and biopharmaceuticals. Here we describe the methods which are effective for creating gene targeting constructs and transgenic moss lines as well as confirming successful homologous integration of the constructs and modification of target gene expression.

Key words

Physcomitrella patens Gene targeting Homologous recombination Knockout construct Protoplast transformation Glyco-engineering Biopharmaceutical production Plant-made pharmaceuticals Molecular farming 

Notes

Acknowledgments

This work was supported by contract research “Glykobiologie/Glykomik” of the Baden-Wuerttemberg Stiftung, by the Excellence Initiative of the German Federal and State Governments (EXC294 to R.R.), and EU-co-funded by INTERREG IV Project A17 “ITP-TIP” (ERDF). We thank Anne Katrin Prowse for proofreading of the manuscript.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Eva L. Decker
    • 1
  • Gertrud Wiedemann
    • 1
    • 2
  • Ralf Reski
    • 1
    • 2
    • 3
    • 4
  1. 1.Plant Biotechnology, Faculty of BiologyUniversity of FreiburgFreiburgGermany
  2. 2.TIP Trinational Institute for Plant Research and Plant Biotechnology, Faculty of BiologyUniversity of FreiburgFreiburgGermany
  3. 3.BIOSS Centre for Biological Signalling StudiesFreiburgGermany
  4. 4.FRIAS Freiburg Institute for Advanced StudiesFreiburgGermany

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