CRISPR-Edited Tobacco Produces Improved Proteins for Pharmaceutical Use

Plants are effective alternative platforms for the production of pharmaceutical proteins. However, differences between plant and mammalian N-linked glycans, such as the presence of β-1,2-xylose and core α-1,3-fucose residues in plants, can affect the effectiveness of plant-derived proteins.

Tobacco is widely used for the expression of recombinant proteins, hence, it is desirable to modify its N-glycosylation machinery to allow the synthesis of complex N-glycans lacking β-1,2-xylose and core α-1,3-fucose. Julia Jansing from Aachen University in Germany and team used multiplex CRISPR-Cas9 to develop tobacco lines without α-1,3-fucosyltransferase and β-1,2-xylosyltransferase activity by inducing mutations on six different genes. The team confirmed the multiple gene knockouts by analysis of endogenous proteins and the recombinant monoclonal antibody 2G12.

The team then compared the CD64‐binding affinity of the 2G12 glycovariants produced in wildtype tobacco, the CRISPR-edited line, and Chinese hamster ovary (CHO) cells. The engineered antibody performed as well as its CHO-produced counterpart.

For more information on this study, read the article in Plant Biotechnology Journal.