C-Man Pathway-based Glyco-engineered Cell Construction Service
Empowering Excellence Through Comprehensive C-Man Pathway-based Glyco-engineered Cell Construction
Glycans on cells are complex and structurally diverse. C-mannosylation is involved in physiological processes such as folding, sorting, and secretion of substrate proteins. CD BioGlyco further generates a library of human embryonic kidney 293 (HEK293) cells with glycosylation capacity by controlling the C-Man glycosylation pathway, which provides a useful tool for analyzing glycan biosynthesis, genetics, and structural characterization. Our researchers provide Glyco-engineered Cell Construction Service by controlling glycosyltransferase genes required for biosynthetic interactions.
CD BioGlyco provides clients with a comprehensive set of glycoengineering cell construction services based on the C-Man type glycosylation.
- Gene editing service
Among the pathway-specific GTf genes are those involved in the initiation of different types of C-mannosylation, including GALNT1-20 (GT27), TMTC1-4 (GT105), DPY19L1-4 (GT98), XYLT1-2 (GT14), and COLGALT1-2 (GT25). We provide both single knockout and double knockout glycosyltransferase gene cell line construction services with the support of a clustered regularly interspaced short palindromic repeats (CRISPR-Cas9)-mediated library. At the same time, we provide efficient knock-in services. In detail, we provide transfection, batch sorting, indel detection by amplicon analysis (IDAA) screening, and sequencing services. - Construction and analysis service
Our researchers use gene editing to engineer and re-express cell line glycosylation capabilities, laying the groundwork for the construction of Cell-based Glycan Display Arrays. We provide stably engineered HEK293 isogenic cell library construction services to generate cells displaying the loss or gain of specific glycosylation features. Glycosylation capacity and role of HEK293 cells assessed by RNA sequencing (RNA-seq) transcriptomics and fluorescence-activated cell sorting (FACS) services.
Fig.1 Schematic diagram of C-Man pathway-based glyco-engineered cell construction service. (CD BioGlyco)
Publication
Paper Title: An atlas of human glycosylation pathways enables display of the human glycome by gene engineered cells
Technology: Gene editing, CRISPR/Cas9, Mass spectrometry
Journal: Molecular Cell
Published: 2019
IF: 16.0
Results: In this article, a library of isogenic HEK293 cells with engineered glycosylation capacity was generated using multiple glycosylation pathways such as C-Man. Combined with gene editing tools and guide RNA (gRNA) libraries, comprehensive gene targeting of glycosylation pathways was facilitated. Targeted control of all human glycosyltransferase genes using knockout or knockin to characterize the C-Man glycosylation pathway to construct a scaleable cell library.
Fig.2 RNAseq analysis of glycosyltransferase genes in HEK293 wildtype cells. (Narimatsu, et al., 2019)
Applications of C-Man Pathway-based Glyco-engineered Cell
- C-Man pathway-based glyco-engineered cells help to support glycan analysis and the exploration of disease biomarkers.
- C-Man pathway-based glyco-engineered cell is suitable for glycan cell engineering to produce specific proteins.
- C-Man pathway-based glyco-engineered cell provides a good basis for the different glycan cell display arrays.
Advantages of Us
- We use the Glycan Display Platform to provide multiple cell line construction services with different glycosylation pathways, such as the Glycolipids (GSL) Pathway, N-linked Glycan Pathway, and Glycan Capping Pathway.
- The CRISPER-Cas9 system offers advantages such as high targeting efficiency and high precision control.
CD BioGlyco has experienced operators and a comprehensive platform to meet the special needs of our clients. We manage and predict glycosyltransferase genes involved in human glycosylation. Please feel free to contact us if you are interested in the C-Man pathway-based glyco-engineered cell construction service.
References
- Narimatsu, Y.; et al. An atlas of human glycosylation pathways enables display of the human glycome by gene engineered cells. Molecular Cell. 2019, 75: 1-14.
- Joshi, H.J.; et al. Glycosyltransferase genes that cause monogenic congenital disorders of glycosylation are distinct from glycosyltransferase genes associated with complex diseases. Glycobiology. 2018, 28(5): 284-294.
