CD Biosynsis offers state-of-the-art Bacillus subtilis Base Editing Services, enabling precise, single-nucleotide substitutions without introducing double-strand breaks (DSBs). Base Editing, utilizing Cas9-derived nickases fused to base deaminases (e.g., Cytidine or Adenine Base Editors), allows for the direct conversion of C>T, G>A, A>G, or T>C bases with high efficiency and minimal off-target effects. This technology is superior to traditional Gene Knock-in for targeted single point mutation, essential for fine-tuning enzyme activity, altering promoter strength (Promoter Engineering), or introducing stop codons for Gene Knockout. We design customized gRNAs and optimize the base editor system for B. subtilis, ensuring rapid, scar-less introduction of precise genetic changes for Pathway Optimization and high-throughput strain improvement.
Get a QuoteBase Editing overcomes many limitations of traditional CRISPR/Cas9 editing, particularly the low efficiency of homology-directed repair (HDR) in non-dividing or industrial microbial strains. In B. subtilis, our Base Editing platform allows for precise, targeted mutagenesis with unprecedented control. This technique is invaluable for applications such as creating subtle amino acid substitutions to improve enzyme thermostability, fine-tuning ribosomal binding site (RBS) efficiency for balanced expression, or introducing premature stop codons to functionally mimic a Gene Knockout. Our designs minimize the window of possible edits, ensuring the desired mutation is achieved with minimal off-target edits, thus accelerating the iterative process of strain engineering.
Cytidine Base Editing (CBE)
Editing of C•G base pairs to T•A base pairs (C>T substitution) for creating missense or nonsense mutations.
Adenine Base Editing (ABE)
Editing of A•T base pairs to G•C base pairs (A>G substitution) for introducing a wider range of functional mutations.
Optimized gRNA Design
Designing gRNAs to position the target base within the optimal editing window of the deaminase domain for maximum efficiency.
Enzyme Activity Fine-Tuning
Introducing non-synonymous mutations to key enzyme residues to adjust specific activity, stability, or substrate binding for Metabolic Pathway Optimization.
Promoter Strength Modulation
Single-base edits within promoter or operator regions to fine-tune transcriptional regulation and achieve balanced gene expression.
Regulatory Site Disruption
Targeted mutations within transcription factor binding sites to disrupt or eliminate native regulatory feedback loops.
Editor Delivery System
Use of specialized, transient B. subtilis shuttle plasmids for the Cas9-deaminase complex to ensure successful editing and easy plasmid curing.
Colony Screening
Initial screening via restriction fragment length polymorphism (RFLP) or high-resolution melt analysis (HRMA) for rapid detection of the mutation.
Deep Sequencing Confirmation
Full Sanger sequencing of the edited locus to confirm the desired single-base substitution and verify the absence of unintended off-target mutations.
A precise, non-DSB approach for single-nucleotide engineering.
Base Editor System Assembly
Transformation & Editing
Screening & Plasmid Curing
Final Sequencing & Delivery
Design: Selection of ABE or CBE; gRNA design to target the desired base within the editing window.
Assembly: Cloning the gRNA cassette into the B. subtilis expression vector containing the base editor machinery.
Transformation: Introduction of the base editor plasmid into the B. subtilis host strain.
Induction: Temporal expression of the base editor system to initiate precise DNA editing.
Screening: High-resolution techniques (HRMA, RFLP) used to identify colonies carrying the desired edit.
Curing: Removal of the base editor plasmid to ensure the genetic change is stable and permanent.
DSB-Free Editing
Avoids error-prone non-homologous end joining (NHEJ) and reduces unintended large deletions or insertions.
High Efficiency & Precision
Achieves highly efficient single-base conversion (C>T, A>G) with minimal bystander or off-target edits.
Fine-Tuning Capability
Ideal for subtle modifications like adjusting enzyme kinetics or modulating Promoter Engineering strength.
Rapid Strain Iteration
Accelerates the DBTL cycle by allowing rapid, multiple sequential base edits in the B. subtilis genome.
"We needed a precise amino acid substitution to enhance our secreted protease stability. Their Base Editing service delivered the exact C>T change without any unwanted mutations, which was impossible with traditional cloning."
Dr. Samuel Liu, R&D Director, Enzyme Engineering
"The ability to rapidly introduce stop codons via Base Editing has become our go-to method for functional Gene Knockout screening in our new Metabolic Pathway Optimization targets."
Ms. Janet Chen, Lead Bioengineer, Synthetic Genomics
"The Base Editing platform allowed us to fine-tune the promoter of a key pathway enzyme, achieving the perfect expression level for balanced flux, a truly scar-less and elegant solution."
Dr. Kenji Tanaka, Principal Scientist, Strain Development
"The turnaround time was much faster than traditional homologous recombination methods. Their robust screening and sequencing confirmed 100% editing accuracy for the critical A>G mutation."
Mr. Alex Johnson, Research Manager, Biologics Research
"We successfully used Base Editing to disrupt a negative feedback loop by targeting a regulatory binding site. The resulting strain showed significantly improved product titer without growth defects."
Dr. Maria Gomez, Group Leader, Industrial Biotechnology
What types of base conversions can you perform in B. subtilis?
We primarily offer Cytidine Base Editing (CBE) for C>T or G>A conversions and Adenine Base Editing (ABE) for A>G or T>C conversions. These cover the most common types of beneficial single-base substitutions.
How does Base Editing compare to traditional homologous recombination (HR) for point mutations?
Base Editing is generally much faster and more efficient than HR for single point mutations, especially in non-dividing B. subtilis cells, because it bypasses the need for an HR template and the double-strand break repair mechanism.
Is Base Editing truly scar-less?
Yes. Since Base Editing only chemically modifies a single base without requiring a DNA repair template, the resulting single-nucleotide substitution is the only change in the genome, making it the ultimate scar-less editing tool.
How do you ensure the Cas9-deaminase plasmid is removed after editing?
We use B. subtilis shuttle plasmids that are temperature-sensitive or easily curable, ensuring the plasmid expressing the editing machinery is cleanly removed post-editing, leaving only the stable genomic base change.
How much does Metabolic Engineering services cost?
The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.
Do your engineered strains meet regulatory standards?
We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).
What to look for when selecting the best gene editing service?
We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.
Does gene editing allow customisability?
Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.
What is the process for keeping data private and confidential?
We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.
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.