O-Linked Glycoengineering Service in Plant Cell

O-Glycosylation in Plant and Mammalian Cells

O-glycosylation in plants and mammals differs not only in the sites of glycan linkage but also in the structure and composition of the glycan. In mammals, O-linked glycoproteins are predominantly mucin-type glycoproteins, GalNAc is added to Ser and Thr residues in the Golgi apparatus. These glycoproteins are further modified with various monosaccharides like GlcNAc, Fuc, Neu5Ac, and Gal, resulting in diverse and complex O-glycan structures. Conversely, in plants, GalNAc O-glycosylation is extremely rare. Ser residues can be glycosylated with a single galactose. The prevalent O-linked glycans of plants are found on Hyp residues which are converted from Pro residues by prolyl-4-hydroxylases (P4H) and are subsequently adorned with arabinose residues. This process leads to the creation of unique plant-specific O-glycan structures. Due to the potential immune response or reduced biological activity derived from non-human O-glycans in terms of plant-based glycoprotein therapeutics production, it is crucial to develop strategies aimed at achieving the humanization of plant O-glycosylation systems.

Fig.1 O-glycosylation in plant and mammalian cells.¹

O-Linked Glycoengineering Services in Plant Cell at Creative Biolabs

Creative Biolabs offers a wealth of expertise and comprehensive services to facilitate O-linked glycoengineering processes in plant-based expression systems. Our goal is to produce glycoproteins with precisely tailored O-glycan profiles that meet the specific needs of our clients. Our team of scientists possesses a deep understanding of the fundamental glycosylation mechanisms that operate in plants. Leveraging this knowledge, we employ cutting-edge techniques in Cellular Genetic Glycoengineering, such as knockout, knockin, and overexpression, to achieve precise glycoengineering outcomes.

Our strategies for O-linked glycoengineering are multifaceted. Firstly, we focus on eliminating undesired native O-glycan modifications present in plant cells. Subsequently, we aim to enhance the glycosylation process by upregulating the expression of glycosyltransferases for mammalian O-glycosylation. These enzymes are crucial for adding specific sugar moieties to glycoproteins in a manner that replicates human-like glycosylation patterns. Therefore, glycoproteins produced in engineered plant systems are characterized by the presence of human-type O-glycans, particularly the prevalent mucin-type glycoproteins.

Plant Cell Glycoengineering for Human Mucin-type Glycoprotein Production

• Removal of plant-specific O-glycosylation
• Eliminating the formation and extension of Hyp residues by knockout P4H genes;
• Preventing the formation and extension of Hyp residues with P4Hs inhibitors.
• Introduction of human mucin-type O-glycosylation
• Overexpression of human GalNAcT2 to initiate the mucin-type O-glycan formation;
• Overexpression of C1GalT1 to elongate the glycan structure;
• Co-expression of UDP-GlcNAc 4-epimerase and UDP-GalNAc transporter to enhance the efficiency of glycosylation initiation;
• Introduction of genes for sialylation pathway to generate sialylated O-glycans.

Advantages of Our Services

• Multiple and comprehensive strategies of glycoengineering
• Efficient and precise genome editing techniques
• High-level expression vector for overexpression
• Customized glycoengineering for tailored glycans

Creative Biolabs is at the forefront of O-linked glycoengineering in plant expression systems. We utilize our expertise, innovative genetic engineering techniques, and a deep understanding of glycosylation processes to deliver customized glycoproteins with the desired O-glycan structures. If you have specific needs related to glycoengineering, please don't hesitate to contact us for further details and tailored solutions.

Reference

1. Schoberer, Jennifer, and Richard Strasser. "Plant glyco-biotechnology." Seminars in Cell & Developmental Biology. Vol. 80. Academic Press, 2018.

Related Services:

1. N-Linked Glycoengineering in Plant Cell