Plants or plant cells can be used to produce pharmacological glycoproteins
such as antibodies or
vaccines. However, these proteins carry N-glycans with plant-typical
residues, which greatly impact the effect or activity of the protein. Two
enzymes are responsible for the addition of plant-specific glycans: ?Â
(1,2)-xylosyltransferase (XylT) and ?Á(1,3)-fucosyltransferase (FucT).
A team of researchers led by S¨?bastien Mercx from Catholic University of
Louvain aimed to knock-out two XylT genes and four FucT genes in Nicotiana
tabacum BY-2 suspension cells using CRISPR-Cas9. Three XylT and six FucT
sgRNAs were designed to target conserved regions. After transformation of N.
tabacum BY-2 cells, genome-edited lines were obtained and their protein
complements were analyzed.
Three lines showed a significant reduction in ?Â(1,2)-xylose and ?Á
(1,3)-fucose, while two lines were completely devoid of them, indicating
inactivation. The KO lines did not show any particular morphological changes
and grew similarly as the wild types.
One KO line was then transformed with genes encoding a human IgG2 antibody.
The IgG2 expression level was as high as in a control transformant which had
not been edited.
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