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.
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.
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.
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.
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)
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)
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.
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