Targeted Gene Knockout

Corynebacterium glutamicum Gene Knockout Service

Custom gene knockout in the industrial GRAS/QPS-qualified host strains (use-specific) Corynebacterium glutamicum.

Custom vector design and construction for your target gene
Homologous recombination or CRISPR-Cas9 editing strategies
PCR and Sanger sequencing validation of knockout clones

Why Choose Our Service?

We deliver verified knockout strains of C. glutamicum with standard QC documentation, enabling you to focus on downstream applications.

Engineered GRAS strain background
Targeted deletion confirmed by sequencing
Scalable from single gene to multiplex projects

GRAS Strain

Custom Strategy

Validated Knockout

Approach

Targeted Gene Knockout in C. glutamicum

We combine established homologous recombination and CRISPR-based methods to generate precise gene deletions in this industrially important amino-acid producer.

Tailored Knockout Design

We design knockout constructs based on your target gene sequence, selecting the most efficient editing strategy—either traditional homologous recombination or CRISPR-Cas9—to evaluate deletion success.

Target gene sequence analysis
SgRNA design (if CRISPR approach)
Donor template with homologous arms

Streamlined Workflow

From vector construction to transformation and clone screening, our workflow is optimized for C. glutamicum to deliver knockout strains in a timely manner.

Vector assembly and quality control
Electroporation or conjugation into C. glutamicum
Colony PCR and Sanger sequencing verification

Custom Strategy

Each project begins with a consultation to define the optimal editing strategy based on your gene and strain background.

Stringent Validation

Knockout is confirmed by PCR and sequencing of the target locus; off-target checks are available upon request.

Reliable Deliverables

You receive verified clonal strains and full QC data, ready for downstream applications such as metabolic engineering or functional studies.

Deliverables

What You Receive

Each knockout project includes the following standard deliverables.

Materials

1–2 verified knockout clones (quantity scoped per project)
Glycerol stocks and/or lyophilized cultures
Transformation control data

Documentation

Vector map and sequence files
PCR and sequencing chromatograms confirming deletion
Project report summarizing design, workflow, and validation results

Project Scope

Customizable Parameters

Project parameters are defined during consultation and recorded in the statement of work.

Parameter	Typical project scope
Target gene	Scoped per project after consultation
Editing strategy	Homologous recombination or CRISPR-Cas9; strategy selected based on gene and strain.
Strain background	C. glutamicum ATCC 13032 or client-provided strain (subject to compatibility).
Number of clones screened	Typically 8–16 colonies screened; additional screening scoped per project.
Off-target analysis	In silico prediction and targeted amplicon sequencing of top off-target sites (scoped).
Antibiotic selection marker	Kanamycin or chloramphenicol resistance cassette; marker removal options available on request.

Workflow

How It Works

Our streamlined process from target design to strain delivery.

Target Evaluation and Strategy Design

We review your target gene sequence, strain background, and project goals to select the most suitable knockout strategy—homologous recombination or CRISPR-Cas9—and design the construct.

Vector Construction

Knockout vectors are assembled with homologous arms or sgRNA/Cas9 components, verified by restriction digestion and sequencing before transformation.

Transformation and Clone Selection

The validated vector is introduced into C. glutamicum via electroporation or conjugation; candidate transformants are selected on antibiotic-containing plates and then screened for the intended edit.

PCR and Sequencing Validation

Candidate clones are screened by colony PCR across the deletion junction, and positive clones are confirmed by Sanger sequencing to support precise gene knockout.

Strain Expansion and Delivery

The validated knockout strain is cultured, quality-controlled, and provided as glycerol stocks or lyophilized pellets along with project documentation.

Applications

Where It Powers Your Research

Knockout strains serve as key tools across multiple C. glutamicum research areas.

Metabolic Engineering

Block competing pathways to redirect carbon flux toward target amino acids or chemicals, improving production yields.

Block competing pathways to increase amino acid or chemical titers
Introduce auxotrophies for selection
Combine with overexpression for flux redirection

Functional Genomics

Investigate gene essentiality, decipher gene function, and construct defined mutant libraries for systems biology.

Investigate gene essentiality and function
Construct defined mutant libraries
Study regulatory networks

Industrial Strain Development

Develop robust production strains with targeted deletions evaluated for effects on substrate utilization, byproduct formation, and tolerance.

Develop robust production strains with targeted deletions
evaluate substrate utilization
Reduce byproduct formation

FAQ

Frequently Asked Questions

Common questions about our C. glutamicum knockout service.

We offer both traditional homologous recombination (using a suicide vector with homologous arms) and CRISPR-Cas9-mediated knockout. The choice depends on target gene characteristics and project timeline; our scientists will recommend the optimal approach during the consultation.
Knockout is confirmed by PCR amplification across the deleted region followed by Sanger sequencing of the amplicon. This assesses the deletion junction and checks for a detectable wild-type allele in the tested amplicon.
Yes, we can accept client-provided strains if they are transformable and compatible with our selection system. A preliminary strain test may be required to support successful transformation.
Turnaround time depends on the complexity of the knockout and the number of clones required. We will provide an estimated timeline during project scoping.

Scientific Literature

References

Yang J, Ma X, Wang X, et al. [Advances in gene editing of Corynebacterium glutamate]. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 2020;36(5):820-828. View on PubMed
Wang Y, Zheng P, Sun J. [Recent advances in developing enabling technologies for Corynebacterium glutamicum metabolic engineering]. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 2021;37(5):1603-1618. View on PubMed
Li C, Swofford CA, Rückert C, et al. Optimizing recombineering in Corynebacterium glutamicum. Biotechnology and bioengineering. 2021;118(6):2255-2264. View on PubMed
Chu LL, Tran CTB, Pham DTK, et al. Metabolic Engineering of Corynebacterium glutamicum for the Production of Flavonoids and Stilbenoids. Molecules (Basel, Switzerland). 2024;29(10). View on PubMed
Zha J, Zhao Z, Xiao Z, et al. Biosystem design of Corynebacterium glutamicum for bioproduction. Current opinion in biotechnology. 2023;79:102870. View on PubMed

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Corynebacterium glutamicum Gene Knockout Service

Why Choose Our Service?

Targeted Gene Knockout in C. glutamicum

Tailored Knockout Design

Streamlined Workflow

Custom Strategy

Stringent Validation

Reliable Deliverables

What You Receive

Materials

Documentation

Customizable Parameters

How It Works

Target Evaluation and Strategy Design

Vector Construction

Transformation and Clone Selection

PCR and Sequencing Validation

Strain Expansion and Delivery

Where It Powers Your Research

Metabolic Engineering

Functional Genomics

Industrial Strain Development

Frequently Asked Questions

References

Ready to Start Your Project?

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