At CD BioGlyco, we recognize that traditional strain selection methods no longer suffice for the precision required in modern biomanufacturing. To bridge this gap, we offer a targeted genome editing service for chassis development, providing the molecular tools necessary to rewire cellular metabolism at the genomic level. By utilizing advanced nuclease systems, we specifically modify, delete, or insert genetic elements within a host's genome to optimize metabolic flux, eliminate competing pathways, and enhance the secretion of target glycoproteins. Whether you are working with microbial systems like Escherichia coli and Saccharomyces cerevisiae, or complex mammalian platforms like Chinese hamster ovary (CHO) cells, our service ensures that your chassis is perfectly tailored to your project's specific requirements. Our goal is to transform standard biological entities into highly efficient, programmable "cell factories" capable of producing high-purity, bio-better therapeutics and industrial enzymes.
Key Technologies
Gene Editing Technology
We utilize proprietary gene editing technology to generate site-specific double-strand breaks (DSBs). By carefully designing the guide RNA (gRNA), we achieve high-efficiency targeting with minimal off-target effects. This method is particularly effective for rapid gene knockouts and large-fragment deletions in a variety of chassis organisms. For projects requiring single-nucleotide resolution without the risks associated with DSBs, we employ base editing and prime editing.
Homology-Directed Repair (HDR)
To achieve stable, site-specific integration of large metabolic pathways or heterologous genes, we harness the cellular HDR pathway. By providing a custom-designed donor template along with the gene editing machinery, we facilitate the seamless insertion of complex genetic circuits into "genomic safe harbors," ensuring stable expression and minimal disruption to the host's fitness.
Targeted Genome Editing Service for Chassis Development
As a core component of our chassis strain development service, CD BioGlyco's targeted genome editing is designed to address the most challenging aspects of host strain optimization. Our service scope covers the entire lifecycle of chassis modification, from initial metabolic mapping to the final validation of the engineered strain. We provide multi-dimensional editing capabilities across a diverse range of host organisms:
Microbial Chassis
Optimization of E. coli, Bacillus subtilis, and various yeast species for the production of human milk oligosaccharides (HMOs) and precursors.
Mammalian Chassis
Sophisticated glycoengineering of CHO and HEK293 cells to ensure human-like N-glycosylation and O-glycosylation patterns in therapeutic antibodies.
Plant and Insect Chassis
Leveraging our chassis development platform to modify Nicotiana benthamiana or Spodoptera frugiperda (Sf9) cells for specialized vaccine and glycoprotein production.
Our implementation strategy involves the systematic removal of "metabolic bottlenecks" that genes that drain resources or produce unwanted glycan byproducts, and the strategic insertion of rate-limiting enzymes. By precisely controlling gene dosage and promoter strength through genomic integration, we provide our clients with a stable, scalable, and patentable chassis.
Workflow
In Silico Design and Strategy Formulation
Using advanced bioinformatics tools, we identify optimal target sites and design high-specificity gRNAs. We perform rigorous off-target prediction to ensure the integrity of the host genome.
Vector Construction and Synthetic Synthesis
We synthesize the necessary vector components, including the cloning of gRNAs into expression vectors, the preparation of ribonucleoproteins (RNPs), and the construction of complex donor templates.
Cellular Transformation and Delivery
The gene editing machinery is delivered into the target chassis using optimized protocols such as electroporation, lipofection, or viral-mediated transduction. For difficult-to-transform strains, we utilize specialized chemical transformation methods.
Selection and High-Throughput Screening
Edited cells are isolated using selectable markers (e.g., antibiotic resistance) or fluorescent reporters. We then employ high-throughput screening (HTS) to identify clones with the desired genetic alterations.
Molecular Validation and Off-Target Analysis
To guarantee accuracy, we perform molecular validation. This includes Sanger sequencing for specific loci and next-generation sequencing (NGS) for whole-genome analysis.
Functional Characterization and Stability Testing
We evaluate the performance of the engineered chassis, including growth kinetics, genetic stability over multiple generations, and the efficiency of target glycan synthesis to ensure the strain is ready for industrial-scale fermentation.
Publication Data
DoI: 10.3390/plants14142100
Journal: Plants
IF: 4.1
Published: 2025
Results: This study demonstrates that the miniature gene editing variant enOsCas12f1 enables efficient targeted genome editing in rice. Codon-optimized enOsCas12f1 recognizes TTN protospacer adjacent motif (PAM) and exhibits DNA cleavage activity in rice protoplasts, with the highest efficiency at TTC PAMs (70.19%). In stably transformed rice, it edits the OsPDS gene at TTC (44.21% efficiency) and TTT (6.12% efficiency) PAM, causing albino phenotypes via 7-29 bp base deletions. Additionally, enOsCas12f1 successfully edits the promoter and 5' UTR of OsDREB1C, achieving up to 64.58% efficiency at the P3 site. Fusion of dead enOsCas12f1 (denOsCas12f1) with the transcriptional activation domain TV enhances OsIPA1 expression by 5.6-8.38 fold. These results establish enOsCas12f1 as a valuable tool for rice genome modification, particularly for generating fragment deletions and gene activation, expanding the plant gene editing toolbox.
Fig.1 Detection of gene editing efficiency in stably transformed rice. (Wang, et al., 2025)
Applications
Therapeutic Glycoprotein Production
Precision editing of mammalian cell lines to eliminate immunogenic glycans (e.g., Neu5Gc or alpha-Gal) and optimize sialylation for extended half-life and enhanced efficacy of monoclonal antibodies (mAbs).
HMO and Rare Sugar Biosynthesis
Engineering microbial chassis to overexpress pathway-specific glycosyltransferases, enabling the cost-effective production of high-purity human milk oligosaccharides for nutraceuticals.
Vaccine Development
Modifying insect or plant chassis to express recombinant viral glycoproteins with specific post-translational modifications (PTMs), ensuring high immunogenicity and safety for next-generation vaccine platforms.
Metabolic Flux Optimization
Rewiring central carbon metabolism in industrial strains to redirect precursors toward glycan synthesis, increasing the yield of hyaluronic acid, heparin, and other glycosaminoglycans (GAGs).
Biofuel and Secondary Metabolite Research
Engineering robust yeast or fungal strains to degrade lignocellulosic biomass and convert it into high-value bioproducts through the introduction of heterologous enzymatic pathways.
Advantages
Unparalleled Genomic Precision
By utilizing our gene editing technology, CD BioGlyco achieves high-efficiency editing even in traditionally recalcitrant species, ensuring that modifications are exactly where you need them with minimal genomic scarring.
Broad Host Compatibility
Our expertise spans the biological kingdoms, offering specialized protocols for bacteria, fungi, plants, insects, and mammalian cells, providing a truly universal chassis development solution for our diverse clientele.
Integrated Glyco-Bioinformatics
We combine genome editing with proprietary metabolic modeling, allowing us to predict the impact of genetic changes on glycan structures before the first experiment is even conducted in the lab.
Rapid Turnaround and Multiplexing
Our streamlined workflow and the ability to perform multiplexed editing (targeting multiple genes simultaneously) reduce the time required to develop a fully optimized production strain.
Frequently Asked Questions
Customer Review
"They successfully eliminated three competing glycan-degrading enzymes in our fungal host, leading to a 5-fold increase in yield. Their technical team's deep understanding of glycobiology was evident throughout the process."
– A.W., Senior Scientist
"We struggled with low-efficiency editing in our proprietary yeast strain for months. CD BioGlyco's gene editing services achieved the desired point mutations in a matter of weeks. The validation data they provided was incredibly thorough and gave us total confidence in our new chassis."
– Q.W., Principal Investigator
"CD BioGlyco's ability to integrate a 12kb pathway into CHO cells using their HDR-based approach was a game-changer for our therapeutic glycoprotein pipeline. The strain's stability has been remarkable."
At CD BioGlyco, our targeted genome editing service is more than just a molecular biology tool; it is a solution for precision bio-design. By combining cutting-edge gene editing technology with deep expertise in glycobiology, we empower researchers to overcome the limitations of natural hosts and build the future of sustainable biomanufacturing. Please feel free to contact us to elevate your chassis development project.
Reference
Wang, J.; et al. Miniature enOsCas12f1 enables targeted genome editing in rice. Plants. 2025, 14(14): 2100. (Open Access)
For research use only. Not intended for any clinical use.
Fig.1 Detection of gene editing efficiency in stably transformed rice. (Wang, et al., 2025)
