O-glycan Core 3 Glycoengineering Service

Importance of O-glycan Core 3

The structure of the core 3 O-glycan is quite concise, consisting of a single N-acetylglucosamine and a single N-acetylgalactosamine unit. The β1-3 N-acetylglucosaminyltransferase (β3Gn-T6) is responsible for catalyzing the core 3 modification (GlcNAcβ1-3GalNAcα-S/T). In mammals, there exists a single β3Gn-T6 enzyme that is predominantly expressed in the stomach, small intestine, and colon. However, in organisms such as Drosophila, the orthologues responsible for synthesizing the core 3 structure have not yet been identified. It is worth noting that core-3 O-glycans have been reported to exert significant suppressive roles in the biology of colorectal tumors. Their impact on tumorigenesis and progression cannot be overlooked, as they play a pivotal role in modulating various cellular processes essential for tumor growth and metastasis. Further investigations are required to unravel the intricate mechanisms through which core-3 O-glycans regulate colorectal tumor biology and to explore their potential implications in therapeutic interventions.

Fig.1 Structure of O-glycan core 3. (CD BioGlyco)

O-glycan Core 3 Glycoengineering Service at CD BioGlyco

• In order to investigate the relationship between tumors and core-3 O-glycan, we establish and characterize three cell lines derived from colorectal tumors that exhibit the expression of C3GnT. By introducing ectopic expression of C3GnT in Cosmc-deficient cells, we are able to eliminate the expression of Tn antigens and promote the synthesis of core-3 O-glycans. That is confirmed by the utilization of the cellular O-glycome reporter/amplification (CORA) technique. It is anticipated that the insights gained from this research will help explore the pivotal role played by C3GnT in colon cancer and develop potential therapeutic interventions.
• In order to investigate the biological role of C3GnT, we produce murine embryonic stem cells that lack the C3GnT gene (C3GnT–/–). The gene-targeting construct currently uses a neomycin selection marker to replace the major part of the coding exon 2 of the C3GnT gene. Due to the absence of an antibody or other molecular probe specific for C3GnT or core 3 O-glycans, we genetically integrate a lacZ reporter immediately after the endogenous C3GnT promoter region to characterize the expression pattern of C3GnT. To measure C3GnT mRNA transcripts, RT-PCR is utilized.

Applications

Advantages

• We have developed three cell lines derived from colorectal tumors that express C3GnT, which can serve as a useful tool for investigating the role of C3GnT in colorectal cancer.
• Our method precisely modifies the C3GnT gene, ensuring the precise deletion or modification of the desired region by employing targeted homologous recombination.
• The incorporation of a neomycin selection marker simplifies the screening process by allowing the identification of cells or organisms that have successfully incorporated the modified C3GnT gene.

CD BioGlyco has earned a well-respected reputation in the field of glycobiology. Our research team consists of highly skilled scientists and researchers who are dedicated to advancing the field. By employing state-of-the-art technologies and novel approaches, CD BioGlyco aims to provide a comprehensive Cell Surface Glycoengineering Service. If you have any questions or would like to obtain further information about their services, please feel free to contact us without hesitation.