CD Biosynsis offers cutting-edge Nannochloropsis spp. Base Editing Services, providing a high-precision, DNA-double-strand-break-free (DSB-free) method for single-nucleotide substitutions. While traditional CRISPR-Cas9 is powerful for creating knockouts via random indels, Base Editing (BE) allows for the direct chemical conversion of one base pair to another (e.g., C:G to T:A or A:T to G:C) at specific genomic coordinates. This technology is uniquely suited for the industrial microalga Nannochloropsis, enabling the precise engineering of lipid-related enzymes, the introduction of herbicide resistance mutations, and the modification of photosynthetic complexes without the genomic toxicity or unpredictable rearrangements often associated with chromosomal breaks.
Our base editing platform addresses the specific genetic requirements of Nannochloropsis, characterized by its compact, haploid genome and robust regulatory landscape. By utilizing catalytically impaired Cas9 (nCas9) fused to specialized deaminase enzymes, we can achieve high-efficiency nucleotide conversion within a narrow "editing window." This capability is revolutionary for algal synthetic biology, as it facilitates "scarless" point mutations that were previously difficult to achieve due to the low efficiency of Homology-Directed Repair (HDR) in this genus. Whether you are performing functional proteomics on the light-harvesting complex or optimizing the catalytic efficiency of enzymes involved in eicosapentaenoic acid (EPA) synthesis, our base editing services provide the precision required for sophisticated algal design.
Get a QuoteBase editing in Nannochloropsis spp. allows for the introduction of specific amino acid changes without altering the surrounding genomic context. This is achieved by targeting a deaminase to a specific DNA site via a guide RNA (gRNA). Once at the target, the deaminase converts the target base (C or A) into a different base, which the cell's replication or repair machinery then fixes into a permanent mutation. This approach is vital for engineering strains with enhanced industrial properties, such as improved tolerance to high light or specialized nutrient utilization.
Our services are powered by codon-optimized base editors specifically designed for the nuclear environment of Nannochloropsis. We account for the microalga's specific transcriptional signals and nuclear localization requirements to ensure that the base editing machinery is expressed at effective levels. By minimizing "bystander" editing—unwanted mutations in adjacent bases—we deliver strains with unprecedented genetic fidelity. This precision enables the systematic study of protein motifs and the rational design of metabolic pathways, moving algal biotechnology from random optimization to predictive engineering.
We provide a range of base editing tools tailored to different types of nucleotide conversions and research objectives in various Nannochloropsis species.
Mechanism
Utilizes a cytidine deaminase to convert Cytosine to Uracil, which is read as Thymine during DNA replication, resulting in a C to T transition.
iSTOP Technology
Enables the induction of premature stop codons (TAG, TAA, TGA) at specific positions, providing a clean method for gene inactivation without the indels of standard CRISPR.
Mechanism
Utilizes an adenosine deaminase to convert Adenine to Inosine, which is read as Guanine, resulting in an A to G transition.
Residue Swapping
Ideal for altering specific amino acid residues in enzymes involved in lipid biosynthesis to modify substrate specificity or catalytic activity.
Herbicide Resistance
Introduction of precise mutations in genes like ALS to confer resistance to specific inhibitors, serving as clean selectable markers.
Regulatory Editing
Editing TATA boxes or transcription factor binding sites to modulate the strength of endogenous promoters without large-scale genomic disruption.
Our rigorous workflow ensures high-purity monoclonal strains with full genomic verification of the target edit.
1. Window Design & Codon Optimization
2. Tool Assembly & Transformation
3. Clonal Isolation & NGS Screening
4. Phenotypic Verification
Identification of the target nucleotide within the optimal editing window. Design of gRNAs and codon optimization of the base editor for the Nannochloropsis nuclear genome.
Construction of algal-specific vectors or preparation of RNP complexes. Delivery via optimized electroporation or biolistic bombardment.
Analysis of protein function, growth kinetics, and metabolic output. Verification that the single-base change has resulted in the desired physiological shift. Delivery of cryopreserved strains.
DSB-Free Integrity
By avoiding double-strand breaks, base editing preserves the overall integrity of the Nannochloropsis genome and prevents unwanted chromosomal rearrangements.
High Efficiency
Our platform is optimized to achieve high-frequency nucleotide conversion, allowing for rapid screening of targeted point mutations.
Scarless Modifications
Enables the creation of precise mutations without leaving behind foreign DNA, ideal for strains intended for industrial applications.
Targeted NGS Proof
We utilize deep sequencing to precisely characterize the editing outcome, ensuring the accuracy of the conversion and monitoring for off-targets.
Technical insights for your Nannochloropsis base editing project.
Contact Us1. How does base editing differ from standard CRISPR-Cas9 in Nannochloropsis?
Standard CRISPR creates double-strand breaks that lead to random insertions or deletions (indels). Base editing chemically converts one base to another without breaking the DNA, allowing for precise single-nucleotide changes.
2. What is the typical "editing window" for these editors?
The activity window is usually a 5-nucleotide stretch (typically positions 4 to 8 relative to the PAM). We design gRNAs to ensure your target base is positioned correctly within this window.
3. Can base editing be used to create gene knockouts?
Yes, by using iSTOP technology to convert a sense codon into a premature stop codon, we can achieve gene inactivation in a more controlled manner than standard knockouts.
4. Is the base edit permanent and stable across generations?
Yes. Once the chemical conversion is fixed by the cell's replication machinery, the mutation is as stable as any other genomic sequence across subsequent generations.
5. Do you offer base editing for chloroplast genes?
Yes, we offer specialized organelle-targeted base editing services utilizing chloroplast-localized base editors to modify photosynthetic complexes directly.
6. How is codon optimization handled for Nannochloropsis?
Diatoms and oleaginous algae have specific codon preferences. We utilize base editor sequences fully optimized for the Nannochloropsis host to ensure maximal expression.
7. Can you perform multiplexed base editing?
Yes, we can deliver multiple gRNAs to target several sites simultaneously, enabling the engineering of multi-site protein domains or complex regulatory regions.
8. What is the typical turnaround time for a base editing project?
A standard base editing project from design to verified monoclonal strain delivery typically takes 14 to 20 weeks.
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.
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
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.