O-GlcNAc Pathway-based Glyco-engineered Cell Construction Service

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)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)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

  1. Hao, X.; et al. Deficient O-GlcNAc glycosylation impairs regulatory T cell differentiation and Notch signaling in autoimmune hepatitis. Front Immunol. 2018, 9: 2089.
  2. 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.
This service is for Research Use Only, not intended for any clinical use.

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