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

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Glyco-Engineering

Part of the book series: Methods in Molecular Biology ((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.

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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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Authors and Affiliations

  1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany

    Eva L. Decker, Gertrud Wiedemann & Ralf Reski

  2. TIP Trinational Institute for Plant Research and Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany

    Gertrud Wiedemann & Ralf Reski

  3. BIOSS Centre for Biological Signalling Studies, Freiburg, Germany

    Ralf Reski

  4. FRIAS Freiburg Institute for Advanced Studies, Freiburg, Germany

    Ralf Reski

Authors
  1. Eva L. Decker
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  2. Gertrud Wiedemann
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  3. Ralf Reski
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Editor information

Editors and Affiliations

  1. University of Natural Resources and Life Sciences, Vienna, Austria

    Alexandra Castilho

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Decker, E.L., Wiedemann, G., Reski, R. (2015). Gene Targeting for Precision Glyco-Engineering: Production of Biopharmaceuticals Devoid of Plant-Typical Glycosylation in Moss Bioreactors. In: Castilho, A. (eds) Glyco-Engineering. Methods in Molecular Biology, vol 1321. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2760-9_15

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  • DOI: https://doi.org/10.1007/978-1-4939-2760-9_15

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2759-3

  • Online ISBN: 978-1-4939-2760-9

  • eBook Packages: Springer Protocols

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