Glycolipids (GSL) Pathway-based Glyco-engineered Cell Construction Service
Empowering Glyco-engineered Cells, One Pathway at a Time
Utilizing glycoengineering to construct cells is an efficient and flexible tool. Depending on the glycosylation pathway, CD BioGlyco provides various types of Glycoengineered Cell Constructs for clients. For example, GSL pathway-based glycan engineering cell construction.
- We utilize the HEK293 cell line as a framework to construct a cell-based glycan display, based on the fact that HEK293 cells have all types of lipid and protein glycosylation capabilities, pathway-specific extensions, and branching features. Moreover, the glycan features produced by GSL are not restricted in HEK293 cells.
- We utilize glycosyltransferase-based clustered regularly interspaced short palindromic repeats (CRISPR) / Cas9 gene editing tools to design and analyze the glycosylation capacity of cell lines. Homozygous cells with specific glycosylation features are constructed by altering glycan complexity branching through knockout (KO) and knock-in (KI) of glycosyltransferase genes.
- We provide predictions of the effects of individual genes on glycan structural features based on the GSL pathway (combinatorial CRISPR/Cas9 KO targeting to control glycosphingolipids results in allelopathic cells differentially displaying glycan features on one or more of these glycoconjugates).
Furthermore, we use a combination of gene editing approaches to eliminate and introduce the ability of cells to glycosylate de novo and develop sub-libraries of stably engineered homozygous cells. On the other hand, we classify glycosyltransferase genes into regulated and non-regulated based on transcriptomic data to further provide differentially regulated glycosylation steps and pathways.
Fig.1 GSL pathway-based glyco-engineered cell construction. (CD BioGlyco)
Publication
Technology: CRISPR–Cas9/CRISPRi tool
Journal: World Journal of Microbiology and Biotechnology
IF: 4.1
Published: 2020
Results: Innovative genome editing technologies based on CRISPR-Cas offer several features and advantages such as high efficiency and specificity as well as ease of handling, which facilitates the construction of cell factories. The CRISPR-Cas system is a three-step antiviral defense mechanism. First, Cas proteins can integrate short DNA spacer regions generated by invaders into CRISPR arrays in the host genome. Second, the CRISPR array consisting of the adapted spacer regions and repetitive sequences is transcribed into a long transcript called preCRISPR RNA (pre-crRNA). At the end of the processing, mature small CRISPR RNAs (crRNAs) are generated from the pre-crRNA and form a double strand with complementary reverse transcription activating RNAs (tracrRNAs). Third, a crRNA-tracrRNA hybridization complex directs Cas9 transferase to the invading DNA target, which acts as a protospacer, leading to the formation of a double-stranded break and the eventual cleavage of the foreign DNA. It is believed that this immune response can destroy the genetic material of the invader and stop the infection through the cleavage of the foreign DNA.
Fig.2 The naturally occurring CRISPR–Cas bacterial immunity. (Hashemi, 2020)
Applications
- Glyco-engineered cell construction can be used to probe the structure and function of human glycoproteins displayed in their natural environment on the cell surface.
- Glyco-engineered cell construction can be used to develop an array of resources for cells and to dissect the biological functions of glycans.
- Glyco-engineered cell construction can be used to research the interactions between glycoproteins and cells, and between cells and cells.
Advantages
- We have advanced gene editing tools and glycoengineering techniques to ensure the stability and reliability of the entire cell construction protocol.
- Our R&D team has been developing novel cell construction technologies and provides clients with Cell-based Glycan Display arrays that are at the forefront of glycobiology.
- We provide our clients with a full range of cell construction, glycan library development, and glycan array development services.
CD BioGlyco provides comprehensive, multiple types of glycoengineered cell constructs. Each of our protocols is quality tested for stability and flexibility to ensure client satisfaction. We hope to be your right-hand man in the field of glycobiology. Please feel free to contact us for more information.
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(2): 394-407.
- Hashemi, A. CRISPR–Cas9/CRISPRi tools for cell factory construction in E. coli. World Journal of Microbiology and Biotechnology. 2020, 36: 96.
