O-GlcNAc Pathway-based Glyco-engineered Cell Construction Service
Unlocking the Power of O-GlcNAc Pathway-based Glyco-engineered Cell
O-GlcNAc glycosylation is a dynamic and inducible post-translational modification. The O-GlcNAc transferase genes responsible for the modification include the O-GlcNAc transferase (OGT) gene and Eogt genes. With our professional Glycan Display platform and efficient gene editing services, CD BioGlyco provides O-GlcNAc pathway-based glycoengineering cell construction services including knockout and knockin of target genes, transfection, expression, and analysis services.
- Reliable gene-editing service
We provide gene editing services for both transcription activator-like effector nuclease (TALEN) and clustered regions of interspersed palindromic repeats (CRISPR/Cas9) systems. Our services include O-GlcNAc glycosyltransferase knockout target design, TALEN plasmid assembly, Cas9/gRNA plasmid construction, and knockout efficiency validation. Our operators have expertise in designing glycosyltransferase gene target regions to ensure functional inactivation. We offer screening and isolation of gene-targeted cells with single- or double-copy inactivation functions.
In addition, we also provide Glyco-engineered Cell Construction services. We provide knockout cells by knockout OGT gene or Eogt gene. Here, both single-knockout cell lines and dual-knockout cell lines, are accomplished in one step by electroporating the relevant vectors into human embryonic kidney 293 (HEK293) cells.
- Transcriptome analysis in HEK293 cell line
Our investigators provide a deep sequencing service for total RNA isolated from HEK293 cells. We provide rapid detection of gene expression profiles for highly expressed genes.
- Sequence validation
For the collected cells, we perform batch sorting for specific expression by fluorescence-activated cell sorting (FACS) service. Furthermore, we screen by indel detection by amplicon analysis (IDAA) and provide Sanger sequencing services for further validation.
- Others
In addition, we provide enzymology and flow cytometry evaluation services.
Fig.1 Schematic diagram of O-GlcNAc pathway-based glyco-engineered cell construction service. (CD BioGlyco)
Publication
Technology: DNA sequencing, Western blot, Flow cytometry
Journal: Frontiers in Immunology
Published: 2018
IF: 7.3
Results: In this study, we established a model of defective O-GlcNAc glycosylation using TALEN-mediated gene-targeted knockdown of the Eogt gene. The results showed that O-GlcNAc glycosylation plays a decisive role in the activation of the cellular Notch signaling pathway. The Eogt gene regulates T cell (Treg) activation and development. Thus, the regulation of the O-GlcNAc pathway is a potential therapeutic target for autoimmune hepatitis.
Fig.2 Knockout of Eogt inhibits Treg differentiation via the NOTCH signaling pathway. (Hao, et al., 2018)
Applications of O-GlcNAc Pathway-based Glyco-engineered Cell Construction
- O-GlcNAc pathway-based glycoengineering cell is the basis of the development and analysis of the Cell-based O-Glycan Array.
- The regulation of the O-GlcNAc pathway is very important for neurological diseases. OGT and O-GlcNAc glycosylation pathways are closely related to the induction of neuropathological phenotypes. It has important significance in the study of neurodegenerative diseases.
- O-GlcNAc glycosylation can be used to regulate the pathogenesis of autoimmune hepatitis. Glyco-engineered cell construction based on the O-GlcNAc pathway is a potential therapeutic and research target.
CD BioGlyco provides multiple glyco-engineered cell construction services based on glycosylation pathways, e.g. N-linked Glycan Pathway, O-GalNAc Pathway, Glycan Elongaton/branching Pathway, and Glycan Capping Pathway. If you have any questions about glyco-engineered cell construction, our scientists and professional technical services team will always help you professionally. Please feel free to contact us for more information.
References
- Hao, X.; et al. Deficient O-GlcNAc glycosylation impairs regulatory T cell differentiation and Notch signaling in autoimmune hepatitis. Front Immunol. 2018, 9: 2089.
- Stolfa, G.; et al. Using CRISPR-Cas9 to quantify the contributions of O-glycans, N-glycans and glycosphingolipids to human leukocyte-endothelium adhesion. Sci Rep. 2016, 6: 30392.
