CD Biosynsis offers high-precision Genome Editing Services for Vibrio natriegens, leveraging the power of the CRISPR-Cas9 system. V. natriegens is rapidly emerging as a superior microbial chassis for synthetic biology due to its exceptionally fast growth rate (doubling time as low as 9.8 minutes). To fully harness this speed for accelerated Design-Build-Test-Learn (DBTL) cycles, high-efficiency, reliable genetic tools are essential. Our service utilizes an optimized CRISPR-Cas9 platform, combined with homologous recombination, to perform precise gene knockouts, targeted knock-ins, and complex pathway modifications. We provide researchers and industries with rationally engineered strains that translate V. natriegens' speed advantage into rapid and scalable bioproduction outcomes.
Get a QuoteThe hyper-fast growth of V. natriegens necessitates genetic tools that can keep pace with its rapid generation time. Traditional, low-efficiency editing methods lead to lengthy selection cycles that negate the host's primary advantage. Our optimized CRISPR-Cas9 system ensures high editing efficiency by maximizing the frequency of targeted double-strand breaks (DSBs) and subsequent homologous recombination (HR). This precision editing is crucial for redirecting the host's native, high-flux metabolism toward the desired product, whether through the complete deletion of a competing pathway or the stable integration of a high-copy biosynthetic cluster. The result is unparalleled acceleration of the entire metabolic engineering process.
Doubling Time
The sub-10 minute doubling time of V. natriegens means strain screening and optimization cycles can be completed significantly faster than with standard hosts like E. coli .
High-Throughput Compatibility
Rapid growth is ideally suited for automated, high-throughput screening (HTS) in microplates, enabling faster library exploration.
Chromosomes I and II
The native dual-chromosome system requires specialized editing strategies, which our platform has optimized to ensure synchronous and stable modification of both replicons for maximum effect .
Multi-Copy Integration
Targeting both chromosomes allows for higher gene dosage and stable genomic integration of pathways without relying on high-copy plasmids.
Aerobic Capacity
Efficient aerobic metabolism provides high precursor availability, making it an excellent host for the production of oxygen-dependent compounds.
Stress Tolerance
Natural tolerance to high salinity and specific stress conditions makes it a robust candidate for industrial fermentation environments.
Our tailored CRISPR platform ensures permanent, precise, and multiplex genomic modifications.
1. Gene Knockout
2. Targeted Knock-in
3. Multiplex Editing
4. Point Mutation (SNP)
High-efficiency, scarless deletion of single or multiple genes (e.g., competing pathways, native degraders) using CRISPR-mediated homologous recombination.
Stable, targeted integration of heterologous biosynthetic pathways or large operons into safe harbor loci on one or both chromosomes.
Simultaneous targeting of up to five or more genes in one cycle using optimized sgRNA arrays, drastically accelerating chassis construction time.
Introducing precise single nucleotide polymorphisms (SNPs) to modify protein function, or optimize regulatory elements like promoters and Ribosome Binding Sites (RBS).
1. Vector Design & Assembly
Design and construction of specialized Cas9 and sgRNA vectors, optimized with V. natriegens-specific promoters and counter-selection markers (e.g., sacB system).
2. High-Efficiency Delivery
Rapid introduction of the editing components (CRISPR plasmid + donor DNA) into the host via optimized electroporation or conjugation protocols, minimizing culture time.
3. Ultra-Rapid Selection
Exploiting the fast growth rate for quick selection/counter-selection of edited colonies. Precise screening to confirm the successful HR event and marker removal.
4. Validation & Curing
Genotype confirmation via sequencing of the edited locus. Final curing step to remove the temporary CRISPR plasmid, resulting in a permanent, marker-free strain.
Scarless and Marker-Free
Final engineered strains are free of any foreign DNA (markers, plasmids), ensuring genetic stability and suitability for regulatory review and large-scale use.
Optimized for Dual Chromosomes
Specific strategies designed to efficiently and accurately target loci on both Chromosome I and Chromosome II simultaneously or sequentially.
Rapid DBTL Cycle Acceleration
High-efficiency editing combined with the host's fast growth rate drastically reduces the time required for each cycle of the metabolic engineering pipeline.
Why is CRISPR-Cas9 essential for V. natriegens?
Due to its extremely fast growth, low-efficiency editing methods (like random mutagenesis) waste valuable time. CRISPR-Cas9 provides the necessary precision and efficiency to keep pace with the host's speed, maximizing the advantage of its short generation time.
How do you handle the dual-chromosome system during editing?
We use specific selection markers and counter-selection strategies, along with optimized homology arms, to ensure that the editing event occurs accurately and is stably maintained on both Chromosome I and Chromosome II, depending on the target locus.
Can you perform multiplex knockouts in V. natriegens?
Yes. Our optimized system allows for the simultaneous targeting of multiple genes on one or both chromosomes using a single CRISPR-Cas9 plasmid expressing an array of guide RNAs.
What kind of validation data do you provide for the edited strain?
We provide full Sanger sequencing of the edited genomic locus, confirmation of plasmid curing, and PCR data verifying the scarless deletion or integration site, ensuring the modification is precise and stable.
What are the typical turnaround times for a single edit?
While the actual editing process is rapid due to the host's fast growth, the entire service (from design to validated strain) typically takes 4-6 weeks, depending on the complexity of the target and the number of editing steps involved.
What culture conditions are required for V. natriegens?
V. natriegens is a halophile, requiring elevated sodium levels (typically 1-3% NaCl) in the media, along with specific nutrient requirements. We use optimized, proprietary media to maximize its growth and transformation efficiency.
Can you integrate large biosynthetic pathways (knock-in)?
Yes. We have developed specialized protocols for the stable genomic knock-in of large DNA fragments (up to 15kb) into the V. natriegens genome, often targeting neutral sites on both chromosomes for increased gene dosage and stability.
Is the final strain guaranteed to be marker-free?
Absolutely. We use counter-selection markers (e.g., sacB) and plasmid curing techniques to ensure that the final delivered strain only contains the desired genomic edit and is free of any antibiotic resistance markers or temporary editing plasmids.
CRISPR-Cas9 technology represents a transformative advancement in gene editing techniques. The main function of the system is to precisely cut DNA sequences by combining guide RNA (gRNA) with the Cas9 protein. This technology became a mainstream genome editing tool quickly after its 2012 introduction because of its efficient, simple and low-cost nature.
The CRISPR gene editing system with its Cas9 version stands as a vital instrument for current biological research. CRISPR technology enables gene knockout (KO) through permanent gene expression blockage achieved by sequence disruption. Various scientific domains including disease modeling and drug screening employ this technology to study gene functions. CRISPR KO technology demonstrates high efficiency and precision but requires confirmation and verification post-implementation because unsatisfactory editing may produce off-target effects or incomplete gene knockouts which impact experimental result reliability. For precise and efficient Gene Editing Services - CD Biosynsis, Biosynsis offers comprehensive solutions tailored to your research needs.
The CRISPR-Cas9 knockout cell line was developed using CRISPR/Cas9 gene editing to allow scientists to remove genes accurately for research on gene function and disease models and pharmaceutical discovery. Genetic research considers this technology essential due to its high efficiency together with simple operation and broad usability.
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CD Biosynsis is a leading customer-focused biotechnology company dedicated to providing high-quality products, comprehensive service packages, and tailored solutions to support and facilitate the applications of synthetic biology in a wide range of areas.