CD Biosynsis offers specialized Nannochloropsis spp. CRISPR-Cas9 Genome Editing Services, providing high-precision genetic modification for this industrial powerhouse of the microalgal world. Nannochloropsis species, including N. oceanica and N. gaditana, are renowned for their extraordinary lipid accumulation capabilities, particularly eicosapentaenoic acid (EPA), making them prime candidates for biofuel and nutraceutical production. Our platform is specifically engineered to overcome the historical challenges of the Nannochloropsis genome, such as the low frequency of homologous recombination and robust epigenetic silencing. By utilizing optimized CRISPR-Cas9 and Cas12a systems, we empower researchers to perform definitive gene knockouts, precise knock-ins, and complex metabolic rewiring.
Our expert team provides end-to-end support for Nannochloropsis engineering, from initial computational target selection to final monoclonal strain validation. We specialize in DNA-free Ribonucleoprotein (RNP) delivery methods, which avoid the integration of foreign DNA and minimize off-target effects, resulting in "clean" edited strains ideal for industrial applications. Whether your goal is to optimize carbon partitioning from starch to lipids, enhance light-harvesting efficiency, or improve stress tolerance in large-scale photobioreactors, our integrated genome editing workflow ensures the delivery of stable, high-performance algal strains with verified genotypes and phenotypes.
Get a QuoteNannochloropsis spp. possess relatively compact genomes (approx. 30 Mb) with high gene density, making them excellent models for systems biology. However, achieving precise genetic control has traditionally been difficult due to the inefficiency of random integration methods. Our CRISPR-Cas9 platform solves these issues by inducing site-specific double-strand breaks (DSBs) that are repaired by the cell's native machinery. We have optimized every component of the system—including codon-optimized Cas nucleases and specialized algal promoters—to ensure robust expression and activity across various Nannochloropsis strains.
Our approach emphasizes the rational redistribution of metabolic flux. By applying CRISPR-mediated editing, we can target competitive pathways, such as those involved in cellulose or starch biosynthesis, to redirect carbon resources toward triacylglycerol (TAG) production. This targeted strategy ensures that engineered strains achieve high specific productivity without compromising growth rates. Our integrated platform also supports the modification of the chloroplast genome, allowing for the direct engineering of the photosynthetic apparatus to improve light utilization and CO2 fixation efficiency in industrial cultivation environments.
We provide a diversified toolkit of genetic modifications tailored to the specific functional requirements of your research or production goals.
NHEJ Indels
Creating frameshift mutations via Non-Homologous End Joining (NHEJ) to achieve permanent loss-of-function for target genes, such as those involved in lipid degradation or competitive carbon sinks.
Dual-gRNA Deletions
Utilizing two gRNAs to excise larger genomic fragments or entire gene clusters, ensuring definitive disruption and easier PCR screening.
Safe Harbor KI
HDR-mediated integration of exogenous metabolic cassettes or reporters into validated genomic safe harbors to ensure stable expression without disrupting essential functions.
Protein Tagging
Precise insertion of fluorescent tags (GFP, mCherry) or affinity markers (FLAG, HA) at the endogenous locus to study protein localization and dynamics.
Pathway Rerouting
Simultaneous targeting of multiple genes in a biosynthetic route using poly-cistronic gRNA arrays, ideal for complex metabolic engineering projects.
Regulatory Tuning
Implementing CRISPRi or CRISPRa for tunable transcriptional repression or activation, allowing for the fine-balancing of metabolic networks.
Our rigorous pipeline is designed to ensure high editing efficiency and the delivery of genetically stable monoclonal strains.
1. Computational Design
2. Tool Assembly & Transformation
3. Monoclonal Screening
4. Verification & Delivery
Selection of target loci and bioinformatic gRNA design with off-target prediction. Full codon optimization of Cas nucleases and reporters for the Nannochloropsis genome.
Preparation of high-purity RNP complexes or specialized algal vectors. Transformation via optimized electroporation or biolistic bombardment.
Genotype verification via Sanger or Next-Generation Sequencing (NGS). Phenotypic validation of metabolic output. Delivery of cryopreserved strains and comprehensive reports.
Biallelic Efficiency
Our platforms are optimized to achieve high-frequency biallelic modifications in Nannochloropsis, ensuring immediate and stable phenotypic expression.
DNA-Free Platforms
Expertise in RNP delivery avoids the integration of foreign DNA, producing "clean" edited strains that are ideal for industrial and regulatory compliance.
Codon Optimization
Proprietary codon-optimization matrices specifically for Nannochloropsis ensure maximal translational throughput of the CRISPR machinery.
Stability Verification
Strains undergo rigorous trials over dozens of generations to ensure that the genetic modifications and phenotypes remain constant over time.
Technical insights for your Nannochloropsis engineering project.
Contact Us1. How do you address gene silencing in Nannochloropsis?
We utilize DNA-free RNP delivery to avoid the triggers of transcriptional silencing or employ vectors with native regulatory elements and optimized codon usage to sustain high expression levels.
2. Can you perform editing in both N. oceanica and N. gaditana?
Yes, we have optimized transformation and editing protocols for the most common industrial Nannochloropsis species and specific lab strains.
3. What is the success rate for biallelic knockouts?
Using our optimized RNP electroporation protocols, we achieve high frequencies of biallelic modifications, which are confirmed through targeted NGS sequencing.
4. How do you ensure the monoclonality of the final strain?
We use automated single-cell isolation (FACS) followed by selective plating. Every delivered strain is verified as monoclonal through genomic junction analysis.
5. Can you target the chloroplast genome specifically?
Yes, we offer specialized chloroplast transformation services using biolistic bombardment to modify the photosynthetic machinery within the plastid genome.
6. Do you offer multiplexed editing for metabolic pathways?
Absolutely. We can deliver multiple gRNAs simultaneously to target redundant genes or several enzymes within a single biosynthetic route.
7. Is the final strain free of antibiotic resistance markers?
If RNP delivery is utilized, the strains are inherently marker-free. For plasmid-based methods, we can use curing strategies to remove markers after editing.
8. What is the typical turnaround time for a knockout project?
A standard single-gene project from design to delivery of a verified monoclonal strain typically takes 14 to 18 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.
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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.