In our project, integrated human practices play a significant part in digging deeper into the issue we are concerned about and achieving our medical benefits and biology goal.

After various interviews, we realize there exists a problem in China that a few people have a comprehensive understanding of the basics of E. coli and its potential applications in disease knowledge and medicine. So, how to build both educational and practical effective products to introduce knowledge to the public and solve clinical problems is also what we aim to figure out in our human practices.

Our product is an efficient gene editing tool, a kit. Aiming at Escherichia Coli Nissle1917, our project intends to construct an efficient genome editing tool based on CRISPR-Cas9 system by increasing the cas9 copy number, and to improve the gene editing efficiency by reducing the escape rate, so as to provide references for the development of other genome editing tools. After obtaining this efficient genome editing tool, it can be applied to the construction of various biofactory chassis cells, and the proportion of target strains obtained can be increased to promote the production and development of various biofactories.

To assess public knowledge about E. coli and related diseases, we hypothesize that there are variations in E. coli awareness among different groups of people, which indicates a need for additional knowledge about E. coli and related diseases. Hypothesizing that teenagers may be in greater need of education of the knowledge, we choose participants aged 18-35 as the compared group to compare and contrast with other groups in both linear regression and logistic regression. We utilized a questionnaire survey comprising 16 questions to test our hypothesis. 452 completed online questionnaires were collected. We determined the total E. coli knowledge scores by tallying the correctness of 8 multiple-choice questions:

1. What type of microbe is E. coli?

2. What are the effects of E. coli on humans?

3. What diseases can E. coli infection cause?

4. What factors contribute to gastrointestinal diseases?

5. How does E. coli infection spread?

6. In which areas is E. coli useful?

7. What dietary precautions can prevent gastrointestinal disorders?

8. How can E. coli infections be prevented in high-risk situations?

 

Out of the 452 participants, 98.09% reside in Mainland China, 1.06% in North America, 0.42% in Singapore, 0.21% in Japan, and 0.21% in Australia. We excluded 4.2% of questionnaires where respondents took less than 60 seconds to complete. The age distribution of valid questionnaires shows that 36.7% are below 18, 37.3% are between 18-36, 32.6% are between 36-56, and 3.5% are above 56 (see Figure 1). Of the participants, 46.9% are students (see Figure 2).

 

 

Figure 1. Ages of valid questionnaire participants

 

 

Figure 2. Professions of valid questionnaire participants

 

The data reveals that 97.9% of participants are familiar with E. coli (see Figure 3), and 67.4% have experienced gastrointestinal diseases, indicating common occurrences (see Figure 4). Regarding dealing with gastrointestinal diseases, 43.2% choose to visit a hospital, 21.5% self-medicate based on their knowledge and experience, 27.9% seek information online, and 7.4% are uncertain (see Figure 5). Notably, most individuals opt for self-treatment, which carries a risk of incorrect medication.

 

 

 

Figure 3. Whether know E. Coli      

 

Figure 4. Whether experience gastrointestinal diseases

 

 

 

Figure 5. Histogram of total scores with the normal curve.

 

To determine if age influences participants' choices for dealing with gastrointestinal diseases, we employed the Pearson Chi-square test due to a sample size exceeding 40, both column and row item counts exceeding 2, all expected values exceeding 1, and expected values between 1 and 5 constituting less than 20%. The null hypothesis, suggesting no age-related effect on choices, was accepted (χ2=10.450, p-value = 0.1315 > 0.05).  

The distribution of total E. coli knowledge scores exhibits a normal curve with a mean of 4.14 and a standard deviation of 1.772, ranging from 0 to 8 (see Figure 6). Most scores cluster around 4.14, showing moderate variability. Some individuals, however, have exceptionally high or low scores.

 

Figure 6. Histogram of total scores with the normal curve

 

Figure 7 illustrates that participants below 18 and above 56 have lower median scores compared to other age groups. Although all groups share the same minimum value and interquartile range (IQR), the 25th percentile (Q1) and 75th percentile (Q3) for participants below 18 and above 56 are lower, as are the maximum values. Using analysis of variance (ANOVA), we confirmed that age differences correlate with differences in total scores. The F value is 5.435, with a p-value of 0.001 < 0.01, supporting this conclusion. For participants aged 18-35 compared to those below 18, the effect on total scores is statistically significant (p-value = 0.001 < 0.01), with an unstandardized B coefficient of 0.688. Similarly, participants aged 36-55 have statistically higher scores than those below 18 (p-value = 0.000 < 0.01), with an unstandardized B coefficient of 0.668. In contrast, there is no statistically significant difference in total scores for participants above 58 years old compared to those below 18.

 

 

 

Figure 7. Box graph of various ages

 

Similarly, Figure 8 displays various total scores based on different professions. Students, service professionals, and liberal professionals have the lowest medians and minimum values, while service workers have a higher maximum score. Other professions have a similar median of 5 correct answers. Administrative assistants and professional staff show similar score distributions with differing outliers. Managers exhibit a narrower range but more outliers. These variations among groups prompted further analysis using ANOVA and linear regression. ANOVA showed a statistically significant difference (F value = 2.224, p-value = 0.031 < 0.05). Subsequently, linear regression revealed that being a professional staff member or administrative assistant results in significantly higher scores than being a student (p-value = 0.004 and 0.03, respectively). The unstandardized B values suggest that professional staff members score on average 0.864 points higher, while administrative assistants score 0.942 points higher than students. In summary, there is a statistically significant difference in scores based on profession, with some professions scoring higher and others exhibiting more variability.

 

 

Figure 8. Box graph of various professions

 

Figure 9 indicates that 96.8% of participants support public education on gastrointestinal and microbiological knowledge. Fisher’s exact test, given that 37.5% of cells have expected counts less than 5 but none less than 1, tested whether age affects support for education. The null hypothesis, implying no age-related effect on support, was accepted as the exact p-value was 0.412 > 0.05. Regarding the form of education, 82.4% of participants believe that organizing campaigns in schools and communities to raise awareness about gastrointestinal health and microbiomes would be beneficial (5.1% "No" and 12.5% "Uncertain") (See Figure 10). Using the Pearson Chi-square test, we found a statistically significant association between age groups and support for these campaigns (χ2 = 19.571, p-value = 0.005 < 0.01), rejecting the null hypothesis. To further analyze this, we employed binary logistic regression, which showed a high goodness of fit (Hosmer-Lemeshow test: p-value > 0.999 and correctness of prediction: 82.4%). Compared to those below 18, the age groups 36-56 and 18-36 had statistically significant tendencies to support the campaigns (p-value = 0.092 < 0.1, p-value = 0.000 < 0.1), with odds ratios of 4.242. In other words, people aged 36-56 are 4.242 times more likely to support these campaigns than those below 18, and those aged 18-36 are 1.658 times more likely.

 

 

 

Figure 9. Whether support public education on gastrointestinal and microbiological knowledge               

 

Figure 10. Whether campaigns in schools and communities are helpful

 

In conclusion, we observed a statistically significant association between age and total E. coli knowledge scores, with individuals below 18 and above 56 tending to score lower. Additionally, there is a statistically significant association between profession and scores, with professional staff and administrative assistants scoring higher than other professions, including students. Most participants have experienced gastrointestinal diseases but tend not to seek medical help or know how to prevent these infections. Therefore, organizing campaigns to educate groups with lower scores, especially teenagers below 18 and students, is essential, which confirms our hypothesis. However, teenagers are not the most supportive of campaigns holding in schools or communities, so alternative educational approaches, such as online campaigns, should be considered.

 

Expert Interview with Professor Li Yanan

We had an interview with Professor Li Yanan from School of Food and Pharmaceutical Engineering, Nanjing Normal University. The following content is the core interview notes and summary.

 

Figure 11. Interview with Professor Li

Stage 1. Current solutions and our competitiveness

In 1917, during World War I, a severe Shigella infection broke out among soldiers on the German front. German microbiologist Alfred Nissle isolated a strain of Escherichia coli from the feces of a soldier who had never been infected with Shigella. He used this bacterium as a medicine to treat other infected soldiers, preventing the spread of the infection. This miraculous strain was later named Nissle1917, abbreviated as EcN. It is the only non-pathogenic strain of E. coli and is considered a probiotic. Made into the drug "Mutaflor" (anti-diarrheal capsule) with EcN as the main ingredient, it has been used in the European market for half a century.

After nearly 20 years of research, the biological characteristics and probiotic functions of EcN have been thoroughly studied. Research has found that EcN not only antagonizes various intestinal pathogens but also regulates the secretion of immune factors in the body, enhancing the host's immune defense capabilities. Clinically, it is mainly used to treat inflammatory gastrointestinal disorders, such as Crohn's disease, ulcerative colitis, and some rarer diseases, such as hemophilia. In recent years, new research results have found that EcN also exhibits its unique anti-tumor characteristics, has tumor-targeting effects, such as melanoma, rectal tumors, and other treatable tumors. When used in combination with chemotherapy drugs, it can enhance the anti-tumor efficacy of the drugs.

Safety Expression: EcN belongs to the serotype O6:K5:H1. Its LPS expresses the semi-rough type O6 antigen. Due to a termination mutation in the polymerase wzy gene on the antigen, its side chain is short and consists of single oligosaccharide repeat units. Its K5 type capsule is serum-sensitive, which means EcN is easily cleared by serum in the body. EcN lacks pathogenic factors commonly found in its pathogenic counterparts and does not carry virulence factors, making it non-pathogenic.

1 Mechanism of Action

1.1 Strengthening the mucosal barrier:EcN can form "cross-talk" between bacteria and cells in the intestinal epithelial cells, thereby enhancing the intestinal mucosal barrier to antagonize pathogens. EcN adheres to intestinal epithelial cells through its F1 type fimbriae and H1 type flagella, continuously secreting bacteriocins locally, thereby inhibiting the adhesion of pathogens to intestinal epithelial cells. EcN can also repair and maintain the integrity of the intestinal epithelium by increasing the expression of tight junction proteins and claudins, indirectly resisting the adhesion and invasion of pathogens.

1.2 Modulating host cell factors: EcN regulates the secretion of immune factors in the body through the TLR-2 and TLR4 dependent pathways, mediates cytokine secretion, and participates in the regulation of the host's immune response, thereby maintaining the homeostasis of the intestine.

1.3 Participating in the regulation of the intestinal flora: EcN can reduce the abundance of Bacteroidetes and increase the abundance of Firmicutes. At the genus level, EcN can increase the abundance of Lactobacillus and reduce the abundance of Bacteroides. The mechanism of action of EcN is that it can colonize the human intestine and prevent pathogens from invading the intestinal mucosa, protecting and repairing the intestinal mucosal barrier.

 

2 Pros and Cons:

2.1 Advantages:

2.1.1 Low toxicity (see 1.1): Numerous animal models and clinical trials have shown that using EcN as a carrier to construct engineered bacteria for drug-targeted delivery and controlled release is safer than taking other therapeutic drugs. It has antagonistic activity against various intestinal pathogens such as Shigella, Vibrio cholerae, Yersinia enterocolitica, Salmonella enteritidis, and invasive E. coli. Moreover, there is a sufficient molecular biology toolbox for EcN engineering.

2.1.2 Tumor targeting: Compared to other drugs, Nissle1917 is a facultative anaerobe, better adapted to the high-pressure environment in the human body. Small molecule drugs, when flowing with the blood to tumor tissues, cannot penetrate due to the density and high pressure inside the tumor, resulting in a low arrival rate. The anaerobic tendency of Nissle1917 enhances its dynamic permeability, allowing it to accumulate in tumor necrotic areas and oxygen-rich areas, with little limitation by factors such as tumor size and the tumor's internal microenvironment. This is the natural advantage of Nissle1917. In the current field of tumor therapy, other treatments are irreplaceable.

2.1.3 Easy to genetically manipulate: The gene sequencing of Nissle1917 has been completed, and its genome and genetic background are relatively clear, making it easier to modify.

2.1.4 Nissle1917 is an extracellular bacterium, easier to clear with antibiotics than intracellular bacteria, offering greater application value.

2.2 Disadvantages:

2.2.1 Application difficulties: Previously, most probiotics on the market used wild-type EcN, initially for the gastrointestinal field. Through research, as an excellent genetic engineering carrier for therapeutic protein expression, its application has significant limitations. Oral administration is relatively safe, but intravenous injection may trigger a series of inflammatory storms to specific proliferation. Therefore, the current method of medication is still very limited.

2.2.2 Instability: In some specific situations, Nissle1917 has limitations in drug expression. For example, when treating certain diseases, proteins and enzymes can be expressed normally with good results. However, the expression of small molecules and chemical drugs is poor, and new chemical synthetic molecules cannot be expressed, making the biopharmaceutical therapy of Nissle1917 still unable to replace existing therapies.

2.2.3 Production is uncontrollable: During the production of EcN, due to high safety requirements, it tends to use constitutive promoters, self-inducing promoters, and conditionally inducing promoters. And the high activity expression of metabolites or recombinant proteins requires plasmids to achieve. However, during the cultivation process, due to the metabolic burden of recombinant plasmids, plasmids are easily lost. It is necessary to add antibiotic resistance genes to maintain plasmid segregation stability, but the use of antibiotics will cause environmental pollution and produce multi-drug-resistant bacteria.

 

Stage 2: Development

As an emerging and efficient treatment technology, at the current stage, EcN, as an exogenous substance expression carrier, is applied to therapeutic protein de-expression therapeutic drugs. When expressed in the body, there is a possibility of instability and low expression. If the CRISPR/CAS9 system is used to knock out endogenous cryptic plasmids and recombine to develop a multi-copy anti-escape system, replacing traditional antibiotics to maintain plasmids, it is a possible experimental application direction in the future. Suppose an engineered bacterium is used to express sialidase. If there is no such system, the sialidase expressed by the plasmid in the body is non-resistant. So in an environment without antibiotics, the plasmid is easily lost, making the bacterium completely ineffective in the body because it no longer expresses therapeutic drugs. So if only this bacterium remains in the body, and this bacterium itself easily triggers an inflammatory response, it will cause certain side effects. Therefore, such a multi-copy anti-escape system was developed to prevent the plasmid from being lost and the protein from being discontinued, ensuring the safety of drug manufacturing and greatly improving the efficiency of drug manufacturing. The professor also expressed strong support for our manufacturing project and expressed a positive view of the future prospects of Nissle1917.

 

 

 

Expert Interview with Dr. Wang Qingzhuo

 

Figure 12. Interview with Dr. Wang

In order to gain insights into the applications, advantages, and principles of CRISPR-Cas, evaluate the project's principles, feasibility, and risks, understand the conversion rate of experimental results into marketable products, we conducted an interview with Dr. Wang Qingzhuo. He holds a Ph.D. from the Key Laboratory of Synthetic Biology at the Institute of Plant Physiology and Ecology, Chinese Academy of Sciences. He is also a postdoctoral researcher at the School of Food and Pharmaceutical Engineering at Nanjing Normal University. Currently, he serves as a lecturer at Nanjing University of Chinese Medicine and is a co-founder of Nanjing Food Qi Biochemical Technology Co., Ltd.

Stage 1. Current technology-Gene editing applications in microbial defense systems

Dr. Wang introduced a microbial immune system named X-line and its application. The immune system is a programmable nuclease that can design on its own cutting position according to the guiding IA sequence, directed to cut the DNA strand or RA strand, thereby degrading foreign DNA or RNA and avoiding viral infection. The scientists applied the system to the gene editing of microorganisms for the treatment of certain genetic diseases.

Stage 2. Application

2.1Application and optimization of gene editing technology

Dr. Wang discussed the application of reducing pottery technology in C editing, including its use in Nissle1917, whether there is a need for reducing pottery in other engineering forces, and how to optimize the cost of products. Dr. Wang pointed out that the application range of reducing ceramic technology is very wide, and there is a certain demand in various fields. For specific user groups, such as scientific research units and pharmaceutical companies, the cost burden is different. In addition, Dr. Wang also discussed reducing the risk of commercial application of Tao Zi technology, and concluded that although there are technical deficiencies and commercial risks, they are generally relatively low.

2.2.Kit optimization design and marketing strategy

Dr. Wang mainly discussed how to optimize the risk of the kit, how to promote the product to the public, how to choose the appropriate price positioning and so on. First, optimization for risk can be adjusted by collecting data and evaluating it; Secondly, the target customers are mainly scientific research institutions and pharmaceutical companies, rather than the general public, so the price positioning needs to consider efficiency and effectiveness. Finally, the consumer group of the product will be different from the existing similar products, and pay more attention to the optimization of gene editing and supporting services.

2.3 Gene editing kit development and sales strategy

Dr. Wang introduced a kit for gene editing, which solves the ceramic particle problem in gene editing and lowers the threshold. The product is inexpensive to produce and does not need to be stored in a refrigerator. The life cycle of this product is difficult to predict, but from the current research, it is expected to become one of the mainstream technologies in the next few years. The kit can be used for the diagnosis and treatment of genetic diseases, such as diseases of the eye. In the promotion process, it is necessary to pay attention to cooperation with manufacturers, publicity and promotion.

2.4.The combination of product promotion and scientific research

Dr. Wang mainly discussed the application and development of a product CRISPR-Cas9 in the treatment of some diseases. The first mentioned that CRISPR-Cas9 can solve some specific diseases such as eye problems, but it needs to be carried out in cooperation with scientific research institutions; Secondly, the research and development direction is to apply CRISPR-Cas9 to E. coli Nissle1917 vector, so that it has a wider range of application scenarios. In the future, with the in-depth study of genetic diseases, the number of treatable diseases will continue to increase, and the application prospect of Chris 9 will be broader.

2.5.Application and exploration of gene editing in Escherichia coli

The practical application and advantages of gene editing technology are introduced. Gene editing technology can be used as a tool to improve the efficiency of gene editing in Nissle1917 reduce escape rates and increase gene editing efficiency. Gene editing in organisms such as E. coli can perform a variety of functions, such as treating enteritis, secreting ketone bodies, or other diseases. By combining the gene editing technology with the method of constant biology, the efficiency and accuracy of gene editing can be improved, and more possibilities can be provided for medical research and application.

1. Vsited TOLOBIO Company

We visited TOLOBIO Company to explore market prospects and enhance the production, transportation, and sales processes of our product.

 

 

Figure 13. Visit to TOLOBIO Company

Firstly, Miss Xiao Wu, the manager of TOLOBIO Company, introduced their product, the 'one-HOur Low-cost Multipurpose highly Efficient System.' ToloBio discovered the highly efficient 'trans-activity' of CRISPR-Cas12 protein and developed the 'HOLMES' nucleic acid rapid detection platform technology based on this unique enzyme, referred to as the 'one-HOur Low-cost Multipurpose highly Efficient System.' Compared to traditional molecular diagnostic techniques, the HOLMES technology based on CRISPR offers advantages such as high sensitivity, high specificity, and rapid results. This technology has been acclaimed as the 'next-generation molecular diagnostic technology' by the American journal 'Science,' holding significant application potential in various fields such as pathogen detection, genetic disease screening, and precision medicine.

 

Secondly, after understanding TOLOBIO Company's product, we conducted an interview with Miss Xiao Wu regarding our questions about the 'one-HOur Low-cost Multipurpose highly Efficient System' and the challenges we face in the production, transportation, and sales of our product.

We categorized and organized Miss Xiao Wu's responses.:

1. Regarding our questions about the 'one-HOur Low-cost Multipurpose highly Efficient System':

1.1 Advantages of the product compared to other brands:

The product's advantages over other brands include:

- Faster and more accurate testing: The product may provide quicker and more precise test results, aiding healthcare institutions and laboratories in making faster and more reliable diagnoses and decisions.

- Portability and ease of use: It may be more portable and user-friendly, suitable for various settings and users, reducing barriers to use.

- Greater potential in multiple fields: The HOLMES technology based on CRISPR may have a broader range of applications, suitable for different types of testing and research.

 

1.2 Potential application areas of HOLMES technology based on CRISPR:

The HOLMES technology based on CRISPR has potential applications in various fields:

- Disease diagnosis and prevention: It may be used for rapid and accurate disease diagnosis, assisting in devising personalized treatment plans.

- Environmental monitoring: It may be used to detect pollutants in water, air, and soil, safeguarding the environment and human health.

- Food safety: It may be used to detect harmful substances or pathogens in food, ensuring food safety.

- Biological research: It may be used in genetic and pathological research, advancing the field of biology.

 

1.3 Iteration cycle of related products:

The iteration cycle of the product typically depends on the speed of technological advancement, market demand, competition, and company strategy. In rapidly evolving and changing technological fields, the iteration cycle may be relatively short, perhaps around 1 to 2 years. However, in some stable markets like traditional healthcare, the iteration cycle may be longer, taking 3 years or more. The iteration cycle aims to continually improve the product, meet market demands, and maintain a competitive edge.

 

2. Regarding questions about our product's production, transportation, and sales processes:

2.1 Future market prospects and industry scale growth:

The gene biotechnology industry has vast market prospects and is expected to experience rapid growth in the coming years. This is primarily due to the increasing focus on personalized medicine and genetic research, as well as continuous technological advancements. Fields such as personalized medicine, gene editing, disease prevention, and diagnostics will drive the development of the gene biotechnology industry. However, market prospects are influenced by various factors such as regulations, ethical considerations, and societal acceptance.

 

2.2 Unexpected expenses in product production:

- Research and development costs: Developing new products or diagnostic tools requires significant funding for research, experimentation, clinical trials, and regulatory approvals.

- Regulatory compliance costs: Compliance with various regulations and standards may require additional costs to ensure product compliance.

- Quality control and assurance: Products must maintain quality, necessitating substantial resources for quality control and assurance.

- Production equipment and technology updates: Ever-changing technology and new equipment may require constant updates.

- Drug registration and approval: Drugs need to be registered and approved in the market, which entails costs and time.

 

2.3 Potential challenges during product implementation:

- Insufficient market acceptance: The product may face challenges of low market acceptance, requiring additional time and resources for promotion and education in the market.

- Difficulty in technological adaptation: Sometimes the product may not be as easy to use in practice as anticipated, requiring user training and technical support.

- Regulatory and compliance issues: Different regions may have varying regulatory and compliance requirements; the product needs to meet these to be successful in those regions.

- Fierce competition: The industry might already have similar products, necessitating innovation or improvement to break through the competitive landscape.

 

2.4 Suggestions for appropriate product pricing:

The appropriateness of product pricing needs to consider various factors:

- Cost analysis: Ensure that the product pricing covers production costs, operating expenses, and a reasonable profit margin.

- Market research: Conduct research on market demand to understand the purchasing willingness and affordability of the target customers, as well as the pricing strategies of competitors.

- Added value: Consider the product's added value and advantages over competitors, and price it to reasonably reflect these advantages.

- Pricing elasticity: Consider the market's sensitivity to prices and adjust prices accordingly to balance market demand and profit.

 

2.5 Suggestions for targeted marketing to the intended customers:

- Choose the appropriate channels: Select suitable marketing channels based on the preferences of the target customers, such as social media, industry exhibitions, professional journals, etc.

- Emphasize unique advantages: Highlight the product's unique advantages and features in marketing to attract the intended customers.

- Regular communication and follow-up: Maintain regular communication with potential customers, understand their needs, address issues promptly, and build good relationships.

 

2.6 Solutions for the storage conditions of high-precision biological reagents during transportation:

When transporting biological reagents with extremely low-temperature requirements, dry ice is typically used. The amount of dry ice required for product transportation can be calculated based on the transport route. Producing dry ice in-house can reduce costs, but a balance needs to be struck between production capacity and cost. Considering a reliable long-term dry ice supplier is also a viable option.

 

Through this visit, we gained insights into cutting-edge products in the gene editing field and also received answers to many of our questions regarding the production, transportation, and sales processes of our product. This has provided us with a clear direction and detailed planning for the company's future development.

 

2. Visited Roche Innovation Center Shanghai: BBD department

 

Figure 14. Visit to Roche Innovation Center Shanghai

We visited the Roche Innovation Center in Shanghai: BBD department to gain insights into their benchmark management practices and to understand their advanced laboratory equipment.

 

Guided by Receptionist XiaoRong Tan, we were given a tour of the Roche Innovation Center Shanghai: BBD department. The first floor of the company consists of public areas, mainly the cafeteria, lounge, and meeting rooms, designed to provide a comfortable environment for employees and enhance their motivation during work.

 

The laboratory areas occupy the second to fourth floors of the building, each designed differently to accommodate laboratories responsible for various research directions, ensuring convenience for researchers during experiments. Moreover, the company has equipped each laboratory with comprehensive facilities. Roche Innovation also maintains clear requirements and processes for laboratory standards, ensuring rigorous conduct of experiments.

 

During the tour, Mrs. Tan introduced us to numerous advanced pieces of equipment, such as The MagNA Pure 96 Instrument, The LightCycler® 96 Instrument, AVENIO Edge System, allowing us to understand the functionality and principles of these devices.

 

Through this visit, we not only gained insights into the importance of employee motivation and well-defined regulations in corporate management but also learned about the pros and cons of different laboratory equipment, providing significant assistance for our future company planning.

 

By visiting companies and engaging in discussions with experts, we gained a deeper understanding of our project's product and the technology behind it. This also helped us clarify the direction for our next steps.

Our target users include those involved in biofactory chassis cell construction, aimed at increasing target strain proportions for biofactory development.

Our efficient gene editing kit targets Escherichia Coli Nissle1917. It enhances CRISPR-Cas9 by increasing cas9 copies, reducing escape rates, serving as a reference for other genome editing tools, and facilitating biofactory chassis cell construction.

We will implement it through online channels like WeChat, academic websites, and collaboration with biology-focused video creators. Offline methods include advertising in biotech park areas, trade show presentations, and direct engagement with potential users in companies and university labs.