Synthetic biology, as an emerging technology, brings various biosafety risks in its research and application that require careful attention. Synthetic biology, being an interdisciplinary field combining biology and engineering, progresses at a much faster pace and larger scale compared to traditional biotechnologies, thereby posing significantly higher potential biosafety risks than other scientific disciplines. Therefore, to ensure that our experiments remain within safe boundaries, we have outlined several precautionary measures. First, we introduce the risks associated with microorganisms and then address some safety concerns based on our innovative perspective. Building on laboratory safety considerations, we provide safety training for each team member before commencing experiments to ensure everyone understands its significance. Through in-depth discussions, we have refined our usage guidelines. To ensure the success of our experiments,

In our interviews with healthcare professionals, we identified potential concerns regarding the misuse and overconsumption of our specialized dietary product by individuals who may not fully understand its proper usage, target audience, and dosages.
To address this, we have implemented the following measures:
Clear Labeling: We will clearly label our product packaging during production, specifying that it is exclusively intended for individuals with phenylketonuria (PKU). Additionally, we will provide specific usage guidelines for different age groups and disease severity levels, along with daily dosage instructions.
Medical-Purpose Registration: We will apply to register our product as a special medical-purpose formulated food, in line with Chinese food safety regulations. This category is designed for individuals with specific dietary needs, ensuring that our product is used appropriately.
Regulated Retail Channels: Special medical-purpose formulated foods are primarily distributed through regulated channels such as hospitals, pharmacies, and specialized health food stores. These outlets are qualified, licensed, and equipped to offer guidance and ensure correct product usage, minimizing the risk of misuse.
By implementing these measures, we aim to prevent potential product misuse among patients.

Furthermore, based on our project, we have identified that numerous iGEM teams and synthetic biology projects may face the risk of potential misuse. Using our project as an example, individuals with phenylketonuria (PKU) could potentially use our project to improve their quality of life. However, if our project were maliciously employed by adding it to the regular diet of healthy infants over an extended period, it could lead to a deficiency of phenylalanine in healthy infants, resulting in a range of health issues. 
It is crucial to draw attention to the risks of misuse associated with synthetic biology projects, as they have the potential to impact individuals and ecosystems negatively. We emphasize the importance of responsible research and application to ensure that such technologies are used for their intended purposes and within ethical boundaries.

To address this concern, we have developed an abuse prevention identification process. By completing this process and answering specific questions, you can assess the level of risk associated with the potential misuse of your project.
"To address this, we have developed a misuse recognition process. Completing this process and answering specific questions can help identify the potential risk of misuse in your project.

Furthermore, we have observed that many teams are focusing on biosafety, particularly in preventing accidental leaks, but there is a lack of in-depth consideration when it comes to the potential misuse of projects. Therefore, we have also created a course PowerPoint presentation on recognizing and preventing misuse in synthetic biology projects. This resource is designed for teams in the field and can be used for broader outreach and education.

"We have created two distinct sty
les of PowerPoint presentations to cater to different age groups. Future iGEM teams can choose the most suitable presentation based on their target audience."

For more advanced practitioners, we have authored a guidebook on preventing misuse in synthetic biology projects. This guidebook is intended to assist seasoned academic professionals in contemplating the risks of misuse within their projects and how to mitigate them."

Gene-edited microorganisms carry the following risks. Firstly, due to antibiotic misuse and the non-specific nature of gene mutations during gene editing, genetically modified bacteria are prone to developing antibiotic resistance, potentially becoming superbugs. The difficulty in eradicating such bacteria makes treatment significantly more challenging than standard bacterial infections. Once these superbugs begin to spread within the population, the consequences could be unimaginable.
Furthermore, gene-edited organisms have unknown impacts on the surrounding ecological systems. Gene editing not only affects the target genes in bacteria but also introduces unpredictable risks related to changes in bacterial behavior and traits. This increases the probability of bacteria gaining a competitive advantage over wild strains. Therefore, if these bacteria were to escape from the laboratory, they would compete with other microbial communities in the local ecosystem, causing disruption, reducing available resources, leading to abnormal species migration, severely disrupting local ecological balance, and diminishing biodiversity.

1: Selection of Chassis Organisms - Choosing Non-pathogenic Chassis
For safety considerations, we selected Escherichia coli 1917 as our chassis organism. Most E. coli strains produce endotoxins that can harm humans if ingested through the final product. Therefore, for safety reasons, we opted for E. coli 1917, which does not produce endotoxins harmful to humans, even if ingested.
2: Gene Selection - Choosing Genes Harmless to Humans/Animals/Plants
For safety throughout the entire process, we used the transport enzyme PheP and phenylalanine ammonia lyase (PAL). Following consultations with experts, we found that both of these enzymes have a wide safety margin and do not pose harm to humans, regardless of their expression levels. Furthermore, even if they undergo mutations (such as increased efficiency or inactivation), they still do not pose safety concerns. So, even if PheP and PAL accidentally leak into the final product and are ingested by patients, they would not harm the human body.
Similarly, the metabolic product trans-cinnamic acid (TCA) and hippuric acid (HA), obtained through human metabolic processes from TCA, are also harmless to the human body, ensuring the safety of the product.


Choice of Application Direction
 1. Application in vivo (Assessing and Evaluating Biosafety Risks in in vivo Applications)
In our choice of application direction, we consider the application of our project within living organisms, specifically within the human body. We understand the importance of assessing and thoroughly evaluating the potential biosafety risks associated with this application.

2. Out-of-body Application for Safety Reasons
In consideration of safety factors and after consultations with safety experts, we have designed our product for out-of-body application, with production taking place in a controlled factory environment. This approach offers several safety advantages:
Legal Recognition: Firstly, out-of-body application products align with legal regulations. We have thoroughly reviewed local Chinese food and drug-related laws and regulations, which clearly state that genetically engineered microorganisms should only be used in products outside the human body and should not be directly applied to the human body or surface. Our product, Antiphe milk powder, undergoes processing within a factory, avoiding direct interaction of engineered microorganisms with the human body, thus complying with legal requirements.
Enhanced Control: Secondly, factory production provides better control over operational processes compared to home preparation, ensuring the safety of the production process and the final product for both human health and the environment. During the production of Antiphe, we take measures to minimize environmental contamination risks. We provide training to factory personnel to prevent bacterial contamination resulting from improper equipment handling. In the process where engineered bacteria are used to produce milk, we employ fermentation tanks with closed systems. When transporting engineered bacteria from the manufacturer to the milk powder processing facility, we convert them into freeze-dried powder to suspend their biological activity, keeping them securely sealed to prevent any potential leakage during transit. Furthermore, to ensure patient safety, we completely remove engineered bacteria from the Antiphe milk powder during processing. Before milk is dried into powder, we employ a bacteria filter with pores smaller than 0.45 micrometers to effectively filter out bacteria, preventing direct contact between engineered bacteria and the patient's body.

We covered various aspects of safety in synthetic biology projects, aligning with the safety requirements of the iGEM competition. Topics included project risk assessment, safety measures, misuse prevention, a misuse recognition process, and the creation of educational materials to promote awareness and safety among different audiences. This comprehensive approach aims to ensure the responsible and secure conduct of synthetic biology research and applications.