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

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

Reliable Service at CD BioGlyco

Proteoglycans (PGs) are important components of the extracellular matrix. It is involved in cell adhesion, growth factor signaling, and inflammatory regulatory processes. PGs consist of a core protein and covalently linked glycosaminoglycan (GAG) chains. O-Xyl pathway is initiated in the Golgi. The biosynthesis of GAG on the cell surface is initiated by O-xylosyltransferase, which transfers xylose to serine residues of the protein. CD BioGlyco provides O-Xyl pathway-based glycoengineering cell construction service through precise gene editing and glycan structure design.

  • The stable genetic engineering strategy

CD BioGlyco provides combinatorial knockout/knockin (KO/KI) glycosylation engineering in mammals by gene regulation. We provide highly stable custom-designed glycosylation-capable cell line construction services. We disrupt endogenous glycosyltransferase genes by KO. All gene editing is verified by sequencing and enzyme-linked immunosorbent assay (ELISA).

In addition, we provide human embryonic kidney 293 (HEK293 cells) culture, construction, and analysis services.

  • Modification service

There is stability and consistency in the de novo expression of the O-Xyl glycosylation pathway. Cell surface glycans are modified at different locations, affecting the biophysical properties and biological functions of the glycans. We provide accurate modification services such as sulfation modification.

Fig.1 Schematic diagram of O-Xyl pathway-based glyco-engineered cell construction service (CD BioGlyco)Fig.1 Schematic diagram of O-Xyl pathway-based glyco-engineered cell construction service (CD BioGlyco)

Publication

Paper Title: Xylosylation of proteins by expression of human xylosyltransferase 2 in plants

Technology: Reverse transcription-PCR, Western blot, Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS)

Journal: Journal of Bioscience and Bioengineering

IF: 3.185

Results: In this article, the recombinant proteoglycan core protein was successfully xylosylated by gene editing regulation of human xylosyltransferase 2. Analysis of the transiently expressed proteins showed that both human xylosyltransferase1 and human xylosyltransferase 2 transfer xylose to the core protein, but the specificity and transfer efficiency of 1 and 2 are different. This study contributes to mammalian proteoglycan molecular biosynthesis.

Fig.2 Enzymatic analysis of xylosyltransferase 1. (Briggs & Hohenester, 2018)Fig.2 Enzymatic analysis of xylosyltransferase 1. (Briggs & Hohenester, 2018)

Applications

  • O-Xyl pathway-based glycoengineering cells are widely used for the recombinant expression and characterizing glycoproteins.
  • O-Xyl pathway-based glycoengineering cells play a huge role in studying the structural properties and physiology of glycans.
  • O-Xyl pathway-based glycoengineering cells lay the foundation for the development of Cell-based Glycan Display Array.

Advantages

  • The gene editing service we provide effectively avoids the stability problem of random plasmid integration.
  • We optimize the coding and construction sites of multiple genes for the targeted KI of codons.
  • The gene editing service we provide stably transcribes during cell culture without affecting the expression of adjacent genes.

CD BioGlyco has rich experience in Glyco-engineered Cell Construction. Moreover, we provide high-throughput Cell-based O-Glycan Array and N-Glycan Array. We have exquisite technology and rich experience, efficiently and accurately complete the test, to ensure that the data is true and reliable. To learn more about the O-Xyl pathway-based glycoengineering cell construction service, please feel free to contact us.

References

  1. Briggs, D.C.; Hohenester, E. Structural basis for the initiation of glycosaminoglycan biosynthesis by human xylosyltransferase 1. Structure. 2018, 26(6): 801-809.
  2. Narimatsu, Y.; et al. An atlas of human glycosylation pathways enables display of the human glycome by gene engineered cells. Mol Cell. 2019, 75(2): 394-407.
  3. Matsuo, K.; Atsumi, G. Xylosylation of proteins by expression of human xylosyltransferase 2 in plants. J Biosci Bioeng. 2018, 126(3): 371-378.
This service is for Research Use Only, not intended for any clinical use.

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CD BioGlyco is a world-class biotechnology company with offices in many countries. Our products and services provide a viable option to what is otherwise available.

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