CD BioGlyco offers a premier gene family editing service for chassis development, designed to provide systemic and precise modifications across multiple genomic loci simultaneously. By leveraging our deep expertise in glycoscience and advanced genome engineering, we enable researchers to bypass the limitations of single-gene manipulation. Whether you are aiming to eliminate competing metabolic pathways, refactor complex glycosylation networks, or enhance the robustness of a production strain, our service provides the technical sophistication required to engineer gene families with unparalleled precision.
Key Technologies
Multiplex Gene Editing Systems
We utilize advanced gene editing systems capable of expressing multiple single-guide RNAs (sgRNAs) from a single polycistronic transcript. This allows for the simultaneous knockout (KO) or knock-in (KI) of 10+ genes within a single family, significantly reducing the timeframe for chassis optimization.
Polycistronic tRNA-gRNA (PTG) Processing
To ensure high expression and precise processing of multiple guides, we employ PTG technology. This system exploits endogenous RNA processing enzymes to release individual functional gRNAs from a single promoter, ensuring uniform and high-efficiency editing across all targeted family members.
Precision Engineering of Genomic Landscapes for Unrivaled Chassis Performance
CD BioGlyco provides a holistic solution for the systematic modification of gene families, covering everything from initial bioinformatic family identification to the final validation of the engineered chassis. The focus of this service is the systemic modification of related genes, such as glycosyltransferases, sugar transporters, or endogenous proteases, that collectively influence the quality and yield of the target product. We provide:
Family Analysis
Identification of all functional paralogs and homologs within the chassis genome to ensure no redundant genes remain to compensate for the desired phenotype.
Customized Editing Strategies
Design of multi-loci strategies including simultaneous deletions, promoter swapping for coordinated regulation, and site-specific large fragment insertions.
Chassis Refactoring
Stripping away non-essential gene families to create a "minimal chassis" that allocates maximum cellular resources toward the target biosynthetic pathway.
Workflow
1. In SilicoDesign and Guide Selection
Our bioinformatics team identifies all members of the target gene family and evaluates their sequence homology. We then design optimized gRNAs with high on-target activity and minimal off-target potential across the entire genome.
2. Multi-cassette Vector Construction
We synthesize and assemble multiplex expression vectors. Depending on the chassis and the number of targets, we utilize either monocistronic or polycistronic architectures to ensure stable and robust expression of the editing machinery.
3. High-Efficiency Transformation and Editing
Using optimized protocols for diverse organisms, including Escherichia coli, Saccharomyces cerevisiae, and CHO cells, we introduce the editing components. Our proprietary methods maximize the frequency of multi-locus modifications in a single transformation event.
4. Clonal Selection and Genotypic Verification
We isolate individual clones and use next-generation sequencing (NGS) or high-throughput polymerase chain reaction (PCR) to verify the editing status of all targeted loci. This step ensures that every gene in the family has been accurately modified.
5. Phenotypic and Multi-omics Characterization
Validated clones undergo rigorous testing to assess growth kinetics, genetic stability, and product yield. We often employ transcriptomics and metabolomics to confirm that the gene family editing has successfully redirected metabolic flux.
6. Final Chassis Delivery and Documentation
The optimized chassis is delivered with a comprehensive technical report, including sequence verification, phenotypic data, and quality control (QC) certificates, ensuring it is ready for immediate integration into your research or production pipeline.
Publication Data
DoI: 10.1104/pp.18.00200
Journal: Plant physiology
IF: 7.7
Published: 2018
Results: This study employs gene editing with a single consensus sgRNA to target the 20-member k1C α-kafirin gene family in sorghum (Sorghum bicolor), aiming to improve grain protein quality and digestibility. The sgRNA targets a conserved N-terminal signal peptide region, inducing small insertions/deletions (InDels) in multiple k1C genes with 92.4% mutation frequency in T1 plants (85/92 individuals). Molecular and biochemical analyses show reduced α-kafirin accumulation, altered protein body morphology, and increased Lys-rich nonkafirins. T2 lines exhibit 1.3-1.5-fold enhanced in vitro pepsin digestibility, 1.8-2.4-fold higher protein-bound Lys, and 2.2-10.6-fold increased free Lys compared to wild-type Tx430. Total protein content was maintained or increased, with some lines showing elevated methionine. This efficient multiplexed gene family editing provides a transgene-removable strategy for sorghum nutritional improvement, addressing key limitations of low Lys content and poor protein digestibility.
Fig.1 Construction of the gene editing system. (Li, et al., 2018)
Applications
Glycoengineering of Biopharmaceuticals
Editing families of glycosyltransferases and glycosidases in mammalian or yeast chassis to produce human-like glycosylation patterns on recombinant proteins, improving therapeutic efficacy and reducing immunogenicity.
Metabolic Flux Optimization
Simultaneously knocking out families of genes involved in competing metabolic pathways (e.g., organic acid production) to maximize the carbon flow toward high-value secondary metabolites and biofuels.
Host Cell Protein (HCP) Reduction
Deleting entire families of endogenous proteases or problematic secretion proteins in production strains to simplify downstream purification and enhance the stability of the final product.
Stress Tolerance Enhancement
Engineering gene families related to cell wall synthesis or oxidative stress response to create a chassis that thrives in the harsh conditions of industrial-scale bioreactors.
Advantages
Unmatched Expertise in Glycobiology
As a leader in glycoscience, CD BioGlyco understands the complex interplay of glycan-related gene families, ensuring scientifically sound project designs.
High-Throughput Multiplexing
We simultaneously target more genes than conventional methods, drastically reducing the number of iterative rounds of editing required to achieve your desired phenotype.
Exceptional Precision and Low Off-target Rates
We utilize proprietary algorithms and advanced variants to ensure that modifications are strictly confined to the target gene family, preserving the overall fitness of the chassis.
Tailored Solutions for Diverse Hosts
Whether your project involves model organisms or non-conventional industrial strains, we have the specialized toolkits and experience to achieve high-efficiency editing across various species.
Frequently Asked Questions
Customer Review
"The gene family editing service provided by CD BioGlyco was instrumental in our effort to refactor the O-glycosylation pathway in our yeast chassis. By knocking out six redundant transferases simultaneously, we saved nearly months of lab work."
– W.Q., Biotechnology Research Institute
"We approached CD BioGlyco to eliminate a family of proteases in our production strain. Their team designed a multiplex gene editing strategy that worked on the first attempt."
– Z.T., Biopharmaceutical Company
"The transition from single-gene engineering to family-wide editing was seamless with CD BioGlyco's help. Their technical team provided invaluable insights into the redundant roles of sugar transporters in our bacterial host."
CD BioGlyco provides the specialized tools, deep scientific expertise, and integrated workflow necessary to execute these complex genomic modifications with speed and precision. Please feel free to contact us for more information on our gene family editing service or to request a customized quote for your chassis development project.
References
Li, A.; et al. Editing of an alpha-kafirin gene family increases, digestibility and protein quality in sorghum. Plant physiology. 2018, 177(4): 1425-1438. (Open Access)
For research use only. Not intended for any clinical use.
Fig.1 Construction of the gene editing system. (Li, et al., 2018)
