Glycogene Editing Service
Glycobiology presents complex research challenges, yet its transformative potential is realized through precise genetic manipulation of glycosylation-controlling genes. CD BioGlyco's glycogene editing services offer a definitive solution, enabling targeted engineering of glycosylation pathways to generate custom cell lines with designed glycan profiles. This approach surpasses traditional glycoengineering limitations, providing a robust and stable methodology for unraveling glycan functionality and advancing next-generation therapeutic development.
Understanding Glycogene Editing
Modifying genes to fundamentally cure diseases is a long-held vision, and the continuous development of gene editing technology is bringing people closer to this aspiration. Gene editing is like a pair of magic scissors that can remove existing DNA and insert replacement DNA to achieve genetic therapy by specifically altering the genetic sequence, destroying toxic genes, and repairing necessary ones.
Fig.1 Gene editing and mechanisms for double-stranded break repair with endogenous DNA. (Li, et al., 2020)
From 1953, when James Watson and Francis Crick discovered the double helix structure of DNA and pioneered the study of modern biology and genetics, to the invention of gene editing tools, gene editing technology has matured from the laboratory to the application. Because of the important role of glycogenes in diseases, glycogene editing occupies an important part of gene editing research. Glycogene editing helps elucidate the glycogenes mechanism that leads to specific glycan types and helps understand the glycosylation process at the system level.
Key Technologies
To accomplish accurate and stable glycogene modifications, CD BioGlyco utilizes state-of-the-art genome-editing technologies. This robust and adaptable platform enables targeted deletion (knockout), insertion (knock-in), or modification of specific glycogenes. These technologies deliver unparalleled specificity and efficiency, empowering researchers to achieve predictable and stable glycoengineering results with minimal off-target impacts.
Precision Glycoengineering: A Paradigm Shift with CD BioGlyco's Glycogene Editing Service
CD BioGlyco provides a flexible and comprehensive service portfolio that goes beyond basic gene editing. We conduct modifications across a diverse range of cell lines, from standard CHO and HEK293 cells to more complex stem cells and primary immune cells. Our expertise accommodates projects of varying complexity—from single-gene knockouts to multiplex editing of multiple glycogenes for complex pathway engineering. We are equipped to manage projects at any scale, whether you need a few modified clones or an extensive collection of engineered cell lines.
Our core knockout solution ensures complete and stable inactivation of glycogene function. Through precise gene editing to disrupt target glycogenes, we generate cell lines deficient in specific glycosylation enzymes—an essential approach for gene function research and the development of simplified glycoforms.
This service enables the targeted insertion of genetic material into the genome. We facilitate the introduction of functional genes to correct mutations, novel genes to establish new glycosylation pathways, or other genetic elements tailored to your research objectives.
For studies requiring partial reduction of glycogene expression, our knockdown service employs RNA interference (RNAi) to suppress gene activity without complete knockout. This method is ideal for investigating the effects of reduced enzyme levels in a subtle, non-lethal manner.
As a specialized knock-in technique, this service allows for the integration of reporter or affinity tags (e.g., fluorescent proteins, His-tags) into glycogenes. This enables visualization, subcellular localization, and purification of protein products, offering critical insights into their functional roles and cellular dynamics.
We introduce extra copies of glycogenes into host cells to significantly upregulate corresponding enzyme expression. This powerful approach enhances glycosylation pathway efficiency and boosts the yield of target glycan structures for various biotechnological applications.
We use the above gene editing methods and strategies to engineer the following glycogenes. Our services include:
We modify genes that encode GT, which are responsible for attaching glucose residues to proteins and lipids. This service is crucial for controlling the synthesis of various glucans and glycoconjugates.
This service focuses on engineering GalT to precisely control the addition of galactose residues. This is vital for applications requiring specific lactose, N-acetyllactosamine, or other galactose-containing structures.
We offer targeted engineering of MTases to regulate mannosylation pathways. This is particularly important for yeast and fungal expression systems and for the synthesis of high-mannose structures in mammalian cells.
This service manipulates the enzymes that add N-acetylglucosamine, a key building block for complex N-glycans. Controlling these enzymes is essential for fine-tuning the branching and structure of N-glycans.
We can engineer GALNT to regulate the initiation of O-glycosylation. This is a powerful tool for controlling the density and type of O-glycans on a target protein.
Our service targets FucT to precisely control the addition of fucose residues to glycans. Fucosylation is critical for blood group antigens and plays a significant role in cell adhesion and inflammatory responses.
This service manipulates ST to control the addition of sialic acid residues. Sialylation is vital for the biological activity and half-life of many therapeutic glycoproteins, and its engineering is a key strategy for improving biopharmaceuticals.
We offer the engineering of UGT to regulate the addition of glucuronic acid. This is particularly relevant for the synthesis of glycosaminoglycans and for studying detoxification pathways.
GAG Polymerase Engineering Service
Our service focuses on modifying GAG polymerases to control the synthesis of long, repeating disaccharide chains that form glycosaminoglycans like heparan sulfate and chondroitin sulfate.
This service targets SULT to precisely control the sulfation patterns of glycans and GAGs. Sulfation is a critical modification that regulates a wide range of biological functions, including receptor binding and cell signaling.
Workflow
Publication Data
Journal: Cancer gene therapy
IF: 5.2
Published: 2022
Results: This study explores ST6GAL1 in breast cancer using in vitro experiments (3D cultures, shRNA knockdown). It finds: knockdown of intrinsic ST6GAL1 reduces breast cancer cell growth, invasiveness, and exosomal ST6GAL1; extracellular ST6GAL1 (from exosomes or recombinant protein) compensates for low intrinsic levels, enhancing proliferation, invasiveness, epithelial to mesenchymal transition (EMT), and cancer stem cells (CSC) markers while suppressing apoptosis; exosomes have uncharacterized cofactors amplifying extracellular ST6GAL1 activity. The work highlights extracellular ST6GAL1's non-cell-autonomous role in breast cancer and its potential as a therapeutic target.
Fig.2 Breast tumor cells released exosome-like vesicles with heterogeneously expressed ST6GAL1. (Hait, et al., 2022)
Applications
- Engineering cell lines to produce glycoproteins with optimized glycosylation patterns significantly enhances the efficacy, stability, and immunogenicity.
- Glycogene editing can be used to engineer cells or viral vectors to express specific glycan antigens, enabling the development of novel vaccines against infectious diseases and cancer.
- Developing cell line models with specific glycogene mutations allows researchers to accurately mimic human genetic diseases, providing a valuable platform for studying disease mechanisms and screening potential drug candidates.
Advantages
- Our use of state-of-the-art gene editing ensures precise targeting of glycogenes, significantly reducing off-target effects.
- From initial project design to final data analysis, our team of seasoned glycobiologists and gene-editing specialists guides you at every step.
- We implement stringent quality control measures throughout the entire workflow, including sequence validation of editing vectors and comprehensive genotyping of final clones, to guarantee the success of your project.
Frequently Asked Questions
As a leader in gene synthesis and gene editing, CD BioGlyco has a lot of practical experience in the field of glycogenomics, providing clients with comprehensive glycogene editing solutions. We also have a strong technical team in Glycogene Delivery, Glycogene Expression Profiling, and Cancer Glycogene Discovery. If you are interested in our services, please feel free to contact us.
Associated Services
After engineering your cells, analyzing the resulting glycan structures is critical. Our mass spectrometry-based glycan analysis services provide in-depth profiling of glycan expression patterns, ensuring your edit had the desired effect.
Our expertise in protein expression systems can complement your engineered cell lines, allowing for the large-scale production of your desired glycoengineered proteins for downstream applications.
To target and study your newly engineered glycans, you may need a specific antibody. Our custom antibody development service can create highly specific antibodies against your unique glycan structures.
References
- Li, H.; et al. Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects. Signal Transduction and Targeted Therapy. 2020, 5: 1. (Open Access)
- Hait, N.C.; et al. Extracellular sialyltransferase st6gal1 in breast tumor cell growth and invasiveness. Cancer gene therapy. 2022, 29(11): 1662-1675. (Open Access)
