biostudies-literatureGoritzer KEuropean Research Council1003065https://www.ebi.ac.uk/biostudies/studies/S-EPMC9493077biostudies-literatureUnknown13Molecular pharming in plants offers exciting possibilities to address global access to modern biologics. However, differences in the <i>N</i>-glycosylation pathway including the presence of β(1,2)-xylose and core α(1,3)-fucose can affect activity, potency and immunogenicity of plant-derived proteins. Successful glycoengineering approaches toward human-like structures with no changes in plant phenotype, growth, or recombinant protein expression levels have been reported for <i>Arabidopsis thaliana</i> and <i>Nicotiana benthamiana</i>. Such engineering of <i>N</i>-glycosylation would also be desirable for <i>Nicotiana tabacum</i>, which remains the crop of choice for recombinant protein pharmaceuticals required at massive scale and for manufacturing technology transfer to less developed counFrontiers in plant scienceEngineering the <i>N</i>-glycosylation pathway of <i>Nicotiana tabacum</i> for molecular pharming using CRISPR/Cas9.PMC9493077774078760331Grandits MFigl RMa JKNavarre CTeh AYGoritzer KGrunwald-Gruber CMercx SfalseEngineering the <i>N</i>-glycosylation pathway of <i>Nicotiana tabacum</i> for molecular pharming using CRISPR/Cas9.Molecular pharming in plants offers exciting possibilities to address global access to modern biologics. However, differences in the <i>N</i>-glycosylation pathway including the presence of β(1,2)-xylose and core α(1,3)-fucose can affect activity, potency and immunogenicity of plant-derived proteins. Successful glycoengineering approaches toward human-like structures with no changes in plant phenotype, growth, or recombinant protein expression levels have been reported for <i>Arabidopsis thaliana</i> and <i>Nicotiana benthamiana</i>. Such engineering of <i>N</i>-glycosylation would also be desirable for <i>Nicotiana tabacum</i>, which remains the crop of choice for recombinant protein pharmaceuticals required at massive scale and for manufacturing technology transfer to less developed coun2022-01-01T00:00:00Z20222025-04-05T12:32:30.053Z2025-04-05T12:32:30.053ZS-EPMC94930773616101010.3389/fpls.2022.1003065