With advances in microbial bioprocessing and genetic engineering, humans have moved beyond traditional fermentation to efficiently produce essential compounds at scale. Through genetic engineering, modified microorganisms can now synthesize previously difficult-to-produce compounds. By optimizing microbial chassis and developing new industrial processes, purer and more cost-effective products are now possible. Today, industrial microorganisms' production capabilities have been revolutionized, with microbial metabolites being used in amino acid production, vaccine and antibiotic development, and isolation of chemicals for organic synthesis.
Creative Biogene applies these advanced genome-editing techniques to improve microbial strains for a variety of applications in the pharmaceutical, environmental, and agricultural fields.
Wide Range of Microorganisms
Creative Biogene supports a vast array of microorganisms, including common industrial strains such as E. coli, S. cerevisiae, and Pichia pastoris, among others.
Scarless Genome Editing
We offer scarless editing, meaning no residual sequences are left behind, ensuring that only the desired genetic modifications are present.
Precise Base-Pair Editing
Our platforms ensure precise editing at the nucleotide level, enabling the highest degree of accuracy in your projects.
Fast and High-Efficiency Service
With a quick turnaround time and highly efficient techniques, we can help meet strict project timelines and reduce development costs.
Flexible Applications
Our services span multiple industries, including industrial applications, pharmaceuticals, environmental management, and agriculture, providing tailored solutions to meet specific needs.
This system allows for gene knockout, site-directed mutation, and gene knock-in, enabling high precision in modifying microbial genomes. CRISPR technology is known for its simplicity, specificity, and low cytotoxicity, making it ideal for a wide variety of microorganisms, including both Gram-negative and Gram-positive bacteria, as well as yeasts.
Using homologous recombination mediated by bacteriophage proteins, recombineering is an excellent tool for gene mutations, deletions, and insertions in microbes. This system is particularly suited for bacterial genome modifications, offering a high level of efficiency and accuracy without introducing foreign DNA sequences.
Pectobacterium carotovorum is a plant pathogen that causes bacterial soft rot in crops, a process regulated by oxygen concentration. DgcO, a soluble globin-coupled sensor protein in P. carotovorum, plays a crucial role in sensing oxygen and modulating the production of c-di-GMP, a bacterial second messenger. The researchers aimed to understand how DgcO influences transcriptional regulation, specifically under aerobic conditions. To achieve this, they deleted the DgcO gene and analyzed transcript levels using RNA sequencing. The analysis revealed that DgcO deletion only impacted transcript levels when the bacteria were grown in oxygen-rich environments. Further investigation showed that the deletion altered the expression of genes involved in metal transport, which was confirmed by inductively coupled plasma-mass spectrometry (ICP-MS) that demonstrated decreased concentrations of essential metals.
Creative Biogene supported this research by constructing the DgcO deletion strain and providing the necessary environment for running ICP-MS assays. These findings highlight the role of DgcO in regulating cellular metal content and oxygen-dependent phenotypes, offering new insights into the molecular mechanisms of P. carotovorum and the potential functions of c-di-GMP in this important phytopathogen.
Figure 1. The researchers measured c-di-GMP levels in wild-type (WT) and ΔdgcO P. carotovorum strains grown under aerobic and anaerobic conditions. They found no significant difference between strains under aerobic conditions, but a notable decrease in c-di-GMP levels in the ΔdgcO strain under anaerobic conditions. (Fekete FJ, et al., 2023)
Carbapenem resistance in Pseudomonas aeruginosa is primarily associated with the regulation of chromosomal resistance factors like AmpC and OprD. The researchers aimed to understand how imipenem enters P. aeruginosa when OprD is absent, and the role of chromosomal β-lactamases in imipenem resistance. They focused on 17 clinical isolates and three laboratory strains, including P. aeruginosa PAO1-derived knock-out mutants. Their investigation revealed that OprD was not essential for imipenem entry and that imipenem susceptibility could be restored by imipenem/relebactam due to the interaction between relebactam and AmpC overexpression induced by imipenem exposure.
To validate these findings, Creative Biogene helped construct the oprD knockout mutant, OprDKOCR, using CRISPR technology. This mutant was essential in confirming the results of both phenotypic and genotypic assays. The researchers utilized RT-qPCR to ensure no unintended polar effects and performed whole-genome sequencing to verify the complete deletion of the gene. The strains were cultured and analyzed in standard growth conditions, with bacterial counts confirmed at the mid-logarithmic phase, providing robust data for understanding the mechanisms behind imipenem resistance and potential therapeutic strategies.
Figure 2. The researchers measured oprD transcript levels and protein production to assess imipenem susceptibility. They performed qRT-PCR and western blot analysis on clinical isolates, using PAO1 and PW2742 as controls. Creative Biogene's CRISPR-generated OprDKOCR strain was crucial for validating the results, ensuring precise knockout, and confirming phenotypic and genotypic observations. (Freed S Jr, et al., 2024)
Whether you need a new strain with enhanced traits or a tailored metabolic pathway to produce a novel compound, Creative Biogene’s expert team is equipped to provide support every step of the way, from strain selection and pathway design to validation and optimization.
If you have any specific requirements or wish to learn more about how our Microbe Genome Editing Services can support your project, feel free to reach out. Creative Biogene is dedicated to helping you achieve your research and production goals with the best possible outcomes. We look forward to collaborating with you on your exciting projects!
Q: What is the difference between CRISPR-Based and Recombineering-Based Microbe Genome Editing?
A: CRISPR/Cas9 System: A precise genetic tool that acts like molecular scissors, allowing scientists to make exact changes in microbial DNA. This system can efficiently add (knock-in) or remove (knockout) specific genes. It works best with microbes that have well-understood CRISPR systems.
Recombineering System: An alternative editing approach that uses the natural process of DNA recombination to modify genes. This method is particularly valuable when:
Q: How long does it take to complete a microbe genome editing project?
A: The timeline for a microbe genome editing project depends on the complexity of the edit and the microbial species. Typically, a simple CRISPR-based knockout may take 3-4 weeks, while more complex modifications (such as multi-gene knock-ins or recombineering edits) may take 6-8 weeks. We also offer expedited services for urgent projects, which may be subject to additional charges.
Q: Do you provide verification services to confirm the success of the genome edits?
A: Yes, every genome editing project includes comprehensive verification steps to confirm the accuracy of the modifications. Verification methods include:
PCR to confirm the presence of the desired mutation
Sequencing to validate the precise change at the genomic level
Phenotypic analysis (if applicable) to confirm functional effects.
We provide detailed reports of all validation steps and results.
Q: Are these genome editing services customizable?
A: Yes, we offer fully customizable genome editing services tailored to your specific research or industrial needs. Whether you require single-gene knockouts, complex multi-gene edits, or pathway modifications, our team can design and execute the optimal editing strategy using CRISPR or recombineering techniques.
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