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- Title
Targeted knockouts ofPhyscomitrellalacking plant-specific immunogenicN-glycans.
- Authors
Koprivova, Anna; Stemmer, Christian; Altmann, Friedrich; Hoffmann, Axel; Kopriva, Stanislav; Gorr, Gilbert; Reski, Ralf; Decker, Eva L.
- Abstract
Using plants as production factories for therapeutic proteins requires modification of theirN-glycosylation pattern because of the immunogenicity of plant-specific sugar residues. In an attempt towards such humanization, we disrupted the genes forα1,3-fucosyltransferase andβ1,2-xylosyltransferase inPhyscomitrella patensby homologous recombination. The singleΔ fuc-tandΔ xyl-tplants, as well as the double knockout, lacked transcripts of the corresponding genes, but did not differ from the wild-type moss in morphology, growth, development, and ability to secrete a recombinant protein, the human vascular endothelial growth factor VEGF121, into the culture medium.N-Glycan analysis, however, revealed the absence of 1,3-fucosyl and/or 1,2-xylosyl residues, respectively. Therefore, the modifications described here represent the key step towards the generation of moss lines suitable for the production of plant-made glycosylated biopharmaceuticals with nonallergenicN-glycans.
- Subjects
PLANT biotechnology; RECOMBINANT proteins; GLYCOSYLATION; PLANT genetic engineering; VASCULAR endothelial growth factors; BIOPHARMACEUTICS
- Publication
Plant Biotechnology Journal, 2004, Vol 2, Issue 6, p517
- ISSN
1467-7644
- Publication type
Article
- DOI
10.1111/j.1467-7652.2004.00100.x