Generation of Arabidopsis thaliana plants with complex N‐glycans lacking β1,2‐linked xylose and core α1,3‐linked fucose

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Abstract
The plant glycosyltransferases, β1,2‐xylosyltransferase (XylT) and core α1,3‐fucosyltransferase (FucT), are responsible for the transfer of β1,2‐linked xylose and core α1,3‐linked fucose residues to glycoprotein N‐glycans. These glycan epitopes are not present in humans and thus may cause immunological responses, which represent a limitation for the therapeutic use of recombinant mammalian glycoproteins produced in transgenic plants. Here we report the genetic modification of the N‐glycosylation pathway in Arabidopsis thaliana plants. Knockout plants were generated with complete deficiency of XylT and FucT. These plants lack antigenic protein‐bound N‐glycans and instead synthesise predominantly structures with two terminal βN‐acetylglucosamine residues (GlcNAc2Man3GlcNAc2).