Genome Engineering of Chlorella vulgaris

1. Strain Characteristics and Biological Background

As one of the most widely used green algae, Chlorella vulgaris features extremely low culture cost, fast growth rate and increasingly mature gene editing technology, making it the most cost-effective strain for industrialized gene editing. Its core applications include: large-scale preparation of medical-grade antioxidant components (such as lutein and phycocyanin), synthesis of raw materials for auxiliary treatment products of neurodegenerative diseases, and expression of proteins for low-cost diagnostic reagents. Meanwhile, it is suitable for scenarios such as technical R&D of small and medium-sized enterprises and project research of scientific research institutions, which can quickly realize technological transformation, balancing cost-effectiveness and practicality.

2. Principles and Processes of Gene Editing Technology

It adopts a low-cost and efficient CRISPR-Cas9 editing system, combined with optimized electroporation transformation technology, which simplifies the operation process, reduces the technical threshold, and meets the needs of industrialization and scientific research popularization. The specific process is as follows:

(1) Target design and sgRNA synthesis: Combined with the genome sequence of Chlorella vulgaris, target sites are accurately designed, and a low-cost sgRNA synthesis scheme is adopted to reduce customer costs;

(2) Simple vector construction: The vector construction process is optimized, and a universal editing vector is used to simplify operation steps, eliminating the need for complex experimental equipment and improving feasibility;

(3) Efficient transformation: The optimized electroporation transformation technology is adopted to improve transformation efficiency, simplify transformation operations, and adapt to the experimental conditions of small and medium-sized enterprises and scientific research institutions;

(4) Positive screening: A convenient resistance screening method is used to quickly screen positive edited strains, reducing screening time and costs;

(5) Editing verification: The accuracy of editing is verified by PCR and sequencing to ensure the editing effect;

(6) Off-target detection (optional): Basic off-target detection services are provided, which can be selected on demand to balance cost and safety;

(7) Large-scale culture: A low-cost culture scheme is optimized to realize rapid large-scale propagation of edited strains, ensuring efficient accumulation of products and adapting to industrialization needs.

Fig.1 Chlorella vulgaris Gene Editing Strategy

3. Service Types

Focusing on high cost-effectiveness and convenience, we provide standardized packages + customized services:

(1) Standardized gene editing packages: Covering single-target knockout, knock-in and point mutation, with prominent cost-effectiveness, suitable for small and medium-sized enterprises and scientific research projects;

(2) Metabolite optimization: Edit related genes to increase the accumulation of medically valuable components such as lutein and phycocyanin;

(3) Low-cost protein expression: Realize low-cost and high-efficiency expression of proteins for diagnostic reagents and medical enzymes;

(4) Research-level strain modification: Provide low-cost edited strains for scientific research institutions to assist in scientific research projects and technical research;

(5)   Industrial culture guidance: Provide low-cost large-scale culture schemes to assist customers in realizing technological landing.

4. Technical Advantages

Relying on the high cost-effectiveness of the strain and GeneRulor optimization, the core advantages meet the actual needs of customers:

(1) Extremely low cost: Simplify the editing process, adopt low-cost reagents and vectors, and the editing cost is reduced by more than 30% compared with similar strains, suitable for small and medium-sized enterprises;

(2) Low technical threshold: No complex experimental equipment is needed, and the standardized process can be quickly mastered, balancing practicality;

(3) Stable editing efficiency: The editing efficiency is more than 75%, with strong stability, which has been verified for a long time;

(4) Fast growth rate: The culture cycle of edited strains is short, which can quickly realize large-scale propagation and reduce production cycle costs;

(5) High cost-effectiveness: Standardized package services balance effect and cost, making it the first choice for scientific research projects and technical R&D of small and medium-sized enterprises;

(6) Strong implementability: The editing system is suitable for industrial culture conditions, which can quickly realize technological transformation without large-scale equipment investment.

5. Project Cycle and Delivery Standards

5.1 Project cycle

(1) Standardized editing package (single target): 40-60 days;

(2) Metabolite optimization: 60-80 days;

(3) Low-cost protein expression: 40-60 days;

(4) Research-level strain modification: 60-80 days.

5.2 Delivery standards:

(1) Physical delivery: Positive edited strains (lyophilized powder + liquid seed solution), large-scale cultures (on demand), target products (on demand);

(2) Technical data delivery: Target design report, vector construction map, transformation and screening records, PCR verification report, sequencing report (off-target detection report provided on demand), low-cost culture manual;

(3) After-sales guarantee: Provide simple operation guidance, answer questions related to strain culture and subsequent application of editing, assist customers in reducing the cost of technological landing, and provide package upgrade support.

References

[1] Kim, J., Chang, K. S., Lee, S., & Jin, E. (2021). Establishment of a Genome Editing Tool Using CRISPR-Cas9 in Chlorella vulgaris UTEX395. International Journal of Molecular Sciences, 22(2), 480. https://doi.org/10.3390/ijms22020480