Genetic Modification of N-Acetylglucosaminyltransferases (GnTs)

GnTs play an important role in glycoprotein biosynthesis. It catalyzes the synthesis of complex glycans with multiple structures. Studies have shown that it mainly affects the synthesis of the O-glycan core structure and the branching of N-linked glycans on glycoproteins. The O-glycan and N-glycan structures are associated with a variety of diseases. GnT gene modification has become a new direction for the treatment of diseases such as diabetes, cancer, etc. In addition, by constructing GnT plant mutants or heterologous expressions of GnT, scientists obtain recombinant proteins with specific structures and functions for disease treatment. GnT gene modification can also be used to study the function of GnT in O-glycan and N-glycan synthesis.

GnT Engineering Service

CD BioGlyco is committed to combining Gene Editing with physiology and case phenotyping to study the function of genes and develop new directions for disease treatment. Our gene modification studies for GnTs involve different models such as mice, plants, and cancer cells.

Fig.1 Different directions of GnT gene modification. (CD BioGlyco)

• Disease-related GnT gene modification service
  O-linked N-acetylglucosaminyltransferase (OGT) is associated with diabetes and its complications. It is mainly responsible for O-GlcNAcylization. We use mouse models to detect changes in diabetic mice by regulating the expression of the OGT gene to develop novel targeted therapies for diabetes.
• GnT gene modification service associated with cancer cells
  GnT-modified N-glycan structures regulate the migration of cancer cells. We construct GnT overexpression or silencing cell lines to detect the effects of cancer cell migration and proliferation, etc.
• GnT gene modification service in plants
  N-glycans play important roles in protein stability, quality control, folding, etc. GnT affects the structure of N-glycans. We establish mutant plants by up-regulating or Down-regulating GnT expression to affect the N-glycan structure of proteins. Ultimately, recombinant proteins with excellent properties are obtained.
• GnT gene heterologous expression service
  Almost all human cell surfaces and secreted glycoproteins are modified by complex N-glycans. To study the function and synthesis of complex N-glycans, we transfer GnT into Escherichia coli for heterologous expression. We will test the enzyme activity and recombinant proteins generated, such as β-1,2-N-acetylglucosaminyltransferase II (hGnT-II) gene.

Published Data

Paper Title: Inhibition of DPAGT1 suppresses HER2 shedding and trastuzumab resistance in human breast cancer

Technology: Gene Overexpression, Gene Silencing

Journal: Journal of Clinical Investigation

IF: 15.9

Published: 2023

Results: Dolichyl-phosphate N-acetylglucosaminyltransferase (DPAGT1) maintains high levels of human epidermal growth factor receptor 2 (HER2) shedding, rendering trastuzumab resistant. DPAGT1 silencing significantly reduced HER2-ECD levels in the culture medium. Targeting DPAGT1 may be a promising strategy for the treatment of trastuzumab-resistant breast cancer.

Fig. 2 Regulatory effects of DPAGT1 on HER2 shedding. (Yang, et al., 2023)

Applications

• Technologies for GnT gene editing are used to study the role of cancer in carcinogenesis, progression, diagnosis, and prognosis.
• Technologies for GnT gene editing have a crucial role in the development of new methods to reduce drug resistance in cancer cells.
• Technologies for GnT gene editing are used for their role in glycan synthesis, which opens up the possibility of obtaining recombinant proteins with structure-specific functions.

Highlights

• We have a professional technical research team and rich experience in molecular cloning. We will construct different target vectors according to client needs.
• We perform gene knockouts, insertions, and other manipulations at different levels. Knockout, overexpression, and other vectors are verified by sequencing as a way to ensure accurate results.
• We have a wide selection of tooling vectors for a variety of expression systems. Various cell lines are constructed.

CD BioGlyco is committed to providing global partners with experimental models rich in polysaccharides and high-level gene editing services. We have a high-level gene modification research team. In addition to GnT, we also provide Galactosyltransferase Engineering Service and N-Acetylgalactosaminyltransferase Engineering Service. If you have any need for gene modification-related experiments, please feel free to contact us.

Reference:

1. Yang, M.; et al. Inhibition of DPAGT1 suppresses HER2 shedding and trastuzumab resistance in human breast cancer. Journal of Clinical Investigation. 2023, 133(14): e164428.