Host Strain Engineering and Phage Development Services
BackgroundService ProcedureOur AdvantagesCase StudyFAQs
Background
Host Strain Engineering
Host strain engineering involves genetically modifying bacterial strains to enhance their desired traits or make them more suitable for specific applications. Improvements can include enhancing metabolic capabilities, increasing resistance to environmental stressors, and modifying interactions with other organisms. The development of genetic engineering tools, such as CRISPR-Cas systems, has significantly advanced this field. These tools enable precise modifications to the bacterial genome, facilitating the development of customized strains for industrial, medical, and environmental applications. The key steps in host strain engineering include selecting the host bacteria, optionally modifying its genome, and validating the engineered host to ensure suitability for the intended application.
Phage Development
Phage development focuses on the study and engineering of bacteriophages—viruses that specifically infect and destroy bacteria. Phages have a natural ability to target and lyse bacterial cells, making them a promising alternative to traditional antibiotics, especially in the face of increasing antibiotic resistance. Modern phage development includes several innovative approaches, such as engineered phages, phage cocktails, phage-encoded enzymes, and phage delivery systems.
Figure 1. Phage gene editing technology in the host bacteria. (A) Phage genome editing through homologous recombination. (B) Differentiation of wild-type and engineered phages using the CRISPR-Cas system. (C) The lambda red recombination system facilitates recombination between editing templates and phage DNA in bacterial cells. (D) Modified retrons generate editing templates via reverse transcription in the host bacterium to facilitate homologous recombination. (E) CRISPR-Cas system utilized for modifying phage genes in host bacteria. (Jia et al., 2023)
Creative BioMart Microbe offers comprehensive host strain engineering and phage development services tailored to your specific objectives. Our multidisciplinary approach integrates advanced genetic engineering, phage biology, and bioprocess optimization to deliver fully characterized microbial and phage platforms suitable for R&D, preclinical development, and biomanufacturing. Contact our team for more information or to request a quote.
Service Procedure
Service Details
Custom Host Strain Engineering
- Receptor Engineering: Enhance phage binding by modifying or overexpressing receptors.
- CRISPR Knockouts: Remove bacterial defenses to improve phage amplification.
- Metabolic Tuning: Adjust growth-related genes for better performance.
- Selectable Markers: Add antibiotic resistance or reporter genes for screening.
Phage Development and Engineering
- Engineered Phages: Tailor-made modifications to enhance lytic activity, host range, or resistance evasion—optionally incorporating CRISPR systems.
- Phage Cocktail Design: Rational formulation of multi-phage blends to target broad-spectrum or resistant bacterial populations.
- Functional Enzymes: Development of phage-derived enzymes (lysins, depolymerases) for direct bacterial lysis or biofilm disruption.
- Stabilization and Delivery: Encapsulation and formulation strategies to increase phage shelf-life, stability, and bioavailability in complex environments.
Our Advantages
- Tailored Genetic Engineering Solutions: Custom strain development based on your application's specific genetic and phenotypic requirements.
- Integrated Phage Development Capabilities: Expertise in both natural and synthetic phage platforms ensures flexibility and innovation in design.
- Rigorous Characterization Protocols: Extensive quality control, including sequencing, imaging, and functional testing, ensures data integrity and reproducibility.
- Scalable and Transferable Processes: Our processes can be adapted for both laboratory-scale research and preclinical or industrial-scale development.
- Collaborative and Transparent Communication: Dedicated project managers provide regular updates, clear documentation, and direct scientific engagement throughout the project lifecycle.
Case Study
Case Study 1: Development of an anti-Acinetobacter baumannii biofilm phage cocktail: genomic adaptation to the host.
The rise of multidrug-resistant (MDR) bacteria like Acinetobacter baumannii has revived interest in phage therapy. In this study, researchers developed an adapted phage, Ab105-2phiΔCI404ad, using evolutionary adaptation from a previously engineered mutant phage. Genome analysis revealed four rearrangements in tail-related genes, expanding the phage's host range by nearly threefold and introducing a depolymerase phenotype in 81% of infected strains.
A phage cocktail combining this adapted phage with depolymerase-producing phage vB_AbaP_B3 showed strong antimicrobial and antibiofilm activity against clinical and reference strains of A. baumannii. Growth curve analyses demonstrated high bactericidal effects, particularly from the adapted phage and the cocktail. However, phage-resistant mutants emerged after 4–5 hours, with resistance being more pronounced in phage B3.
Figure 2. Infection curves. (A) Infection curves for phages vB_AbaP_B3, Ab105-2phiΔCI404ad, and the cocktail composed by both phages, in a planktonic culture of the strain Ab404_GEIH-2010 at MOI 0.1, 1, and 10. (B) Infection curves for phages vB_AbaP_B3, Ab105-2phiΔCI404ad, and the cocktail composed by both phages, at MOI 10 in planktonic culture of 6 strains of A. baumannii. (Blasco et al., 2022)
Case Study 2: Phage-antibiotic combinations against multidrug-resistant Pseudomonas aeruginosa in in vitro static and dynamic biofilm models.
This study evaluated the use of phage-antibiotic combinations (PACs) as a strategy for treating multidrug-resistant Pseudomonas aeruginosa infections. Ten multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains and five phages were characterized, with two key strains (AR351 and I0003-1) used in detailed analyses. Biofilm time-kill assays revealed that PACs showed synergistic or bactericidal activity.
Exposure to PACs modestly increased ciprofloxacin susceptibility in both strains. An optimized three-phage cocktail (EM + EC + 109) with ciprofloxacin showed the most effective biofilm reduction. Notably, phage 109 prevented resistance development to the other phages within the cocktail. These results highlight the potential of PACs to enhance antimicrobial efficacy and prevent resistance in biofilm-associated MDR/XDR P. aeruginosa infections.
Figure 3. Biofilm model for AR351 versus phage cocktail ± ciprofloxacin. In vitro PK/PD biofilm model results for all phage and CIP combinations against P. aeruginosa strain AR351. (Holger et al., 2023)
FAQs
Q: Which bacterial strains can be engineered through your service?
A: We support both Gram-negative and Gram-positive bacteria, including Escherichia coli, Pseudomonas spp., Staphylococcus spp., Bacillus spp., and Lactobacillus spp. Custom strain development is also available upon request.
Q: Can you work with anaerobic bacteria?
A: Yes. We have experience isolating phages and engineering strains under anaerobic conditions, including Clostridium and Bacteroides species.
Q: Do you provide synthetic or engineered phage services?
A: Yes. We offer synthetic biology approaches, including de novo genome synthesis, modular engineering, and recombination-based modification of phage genomes.
Q: Is there a minimum project size or scope?
A: We accommodate projects ranging from single strain or phage developments to comprehensive platform builds. We tailor each engagement to client needs and scale.
Q: Can you customize phage cocktails for complex bacterial infections?
A: Absolutely. We specialize in developing customized phage cocktails tailored to target specific bacterial pathogens. Our team of experts uses advanced techniques to identify and combine multiple phages that work synergistically to combat infections.
Q: How do you ensure the phages developed are safe and effective for my application?
A: Our Phage Development Services provide targeted, safe, and effective phage solutions. We rigorously screen, test, and validate phages for specificity, purity, and stability.
References:
- Blasco L, Bleriot I, González De Aledo M, et al. Development of an anti-Acinetobacter baumannii biofilm phage cocktail: genomic adaptation to the host. Antimicrob Agents Chemother. 2022;66(3):e01923-21. doi:10.1128/aac.01923-21
- Holger DJ, El Ghali A, Bhutani N, et al. Phage-antibiotic combinations against multidrug-resistant Pseudomonas aeruginosa in in vitro static and dynamic biofilm models. Silverman JA, ed. Antimicrob Agents Chemother. 2023;67(11):e00578-23. doi:10.1128/aac.00578-23
- Jia HJ, Jia PP, Yin S, Bu LK, Yang G, Pei DS. Engineering bacteriophages for enhanced host range and efficacy: insights from bacteriophage-bacteria interactions. Front Microbiol. 2023;14:1172635. doi:10.3389/fmicb.2023.1172635
