GPI Anchor Pathway-based Glyco-engineered Cell Construction Service
Unlocking the Power of Glyco-engineering with GPI Anchor Pathway Precision
Glycosylphosphatidylinositol (GPI) anchoring is located at the C-terminus of many eukaryotic proteins and is a post-translational modification. GPI anchor pathways are often tightly linked to certain physiological activities of cells. Examples include signal transduction, targeting the apical membrane, and more. At CD BioGlyco, we have constructed a Glycan Display Platform, and combine GPI anchoring with Cell-based Glycan Display techniques to provide GPI anchor pathway-based glyco-engineered cell construction services.
- We chose the HEK293 cell line, which is suitable for a large number of glycoproteins, as a platform for the demonstration of glycans for cell construction.
- We utilize the GPI pathway to anchor attached proteins in the outer leaflet of the cell membrane (GPI-anchored proteins are transferred to the cell membrane in vitro and in vivo), causing cells to undergo glycosylation. At the same time, we use RNA sequencing (RNA-seq) transcriptomics to predict the glycosylation capacity of HEK293 cells.
In this process, we use chemical synthesis to provide a pathway for GPI-anchored proteins. - We add and remove glycan complexity branches by KO and/or KI of the glycosyltransferase genes to generate homozygous cells showing loss or gain of specific glycosylation features. We use the property that cellular glycans can undergo homologous recombination with exogenous homologous sequences to inactivate specific glycosyltransferase target genes. Alternatively, one gene can be replaced with another by homologous recombination to generate homozygous cells.
Fig.1 GPI anchor pathway-based glyco-engineered cell construction service. (CD BioGlyco)
Publication
Technology: Cellular constructs, gene coding
Journal: Applied Microbiology and Biotechnology
IF: 5
Published: 2008
Results: A new system of cell surface display in Aspergillus oryzae (A. oryzae) was developed by the authors. First, the authors cloned five genes encoding putative cell wall localization proteins in the A. oryzae genome and examined the function of these cell surface GPI anchors by fusing them to the C-terminus of green fluorescent protein (GFP). In addition, the authors used MP1 and CWP proteins as anchor proteins, and the cell surface of recombinant A. oryzae was strongly stimulated by GFP. Enzyme activity was observed on the surface of the cells when these proteins had been applied as anchor proteins to demonstrate A. oryzae glucosidase. In particular, the β-glucosidase activity of recombinant A. oryzae using MP1, a GPI-anchoring protein, was higher than that of CWP. Based on these results, it can be concluded that the MP1 proteins can be used as GPI-anchoring proteins in A. oryzae.
Fig.2 Observation of A. oryzae by fluorescent microscopy. (Adachi, et al., 2008)
Applications
- Based on the GPI-anchored pathway, glycoengineering techniques can be used to develop membrane-anchored devices for cell surface glycoproteins.
- Glyco-engineered Cell Construction techniques can be used to probe the biological functions of cells. Examples include intercellular signaling, cell differentiation, and cell proliferation, among others.
- Glyco-engineered cell constructs can be used to probe cell surface glycan epitopes for the identification of their natural ligands.
Advantages
- We have a professional R&D team and our R&D personnel are well versed in the cell construction process. They have rich practical experience and provide high-quality cell construction services.
- Our world-leading glycoengineering technique ensures the stability and reliability of our glycoengineered cell construction services.
- Our glycoengineered cell construction services based on GPI-anchored pathways apply to a wide range of industries, including glycobiology, life sciences research, and so on.
CD BioGlyco has a unique glycan display platform that provides clients with a wide variety of cell-based glycan display services. For example, constructing cells using glycoengineering techniques, and constructing cell-based glycan arrays, etc. If you need our quality services, please feel free to contact us for details.
References
- Adachi, T.; et al. Construction of an Aspergillus oryzae cell-surface display system using a putative GPI-anchored protein. Applied Microbiology and Biotechnology. 2008, 81: 711-719.
- Paulick, M.G.; Bertozzi, C.R. The glycosylphosphatidylinositol anchor: a complex membrane-anchoring structure for proteins. Biochemistry. 2008, 47(27): 6991-7000.
- Uhler, R.; et al. Glyco-engineered HEK 293-F cell lines for the production of therapeutic glycoproteins with human N-glycosylation and improved pharmacokinetics. Glycobiology. 2021, 31(7): 859-872.
