Design Safety
We worked exclusively with Biosafety Level 1 organisms, including Escherichia coli DH5-α, and Lactobacillus plantarum L168. In addition, the Parts in our project were all Biosafety Level 1 Parts as well.
System 1
In system one, we chose Lactobacillus plantarum L168, a common probiotic, as the chassis, and PLDHLH673 as the vector.The engineered bacteria boost ferrichochrome expression by overexpressing the key enzyme gene.
For biosafety, we designed a suicide switch that can stop them from overgrowing inside the human body and prevent their release into the environment. The suicide switch is activated by a transferable nisin-controlled expression (NICE) system based on the combination of the nisA promoter and nisRK regulatory gene.
System 2
In System two, we also chose Escherichia coli DH5-α, a common probiotic, as the chassis, and PLDHLH673 as the vector. The engineered bacteria boost protoporphyrin IX expression by overexpressing the hemA gene.
For biosafety, we designed the same killing switch, which was activated by a transferable nisin-controlled expression (NICE) system based on the combination of the nisA promoter and nisRK regulatory gene.
It should be noted that our engineered bacteria are currently only grown in the laboratory and will not be released into the environment.
Lab safety
Lab Condition
We work in a neat and safe Lab under professional supervision,which is a Level 1 Standard Microbiological Lab. The lab possesses standard laboratory equipment, and we carried out all our experiments in proper places. Additionally, instructions and warning signs can be seen everywhere in the lab,every team member have received safety training.We also applied a certain number of dedicated rules to ensure Lab safety.
During experiment,we use a biosafety cabinet to handle biological materials,and only use non-pathogenic strains of low-risk organisms to ensure that no hazardous micro-organisms will flow into the natural environment.
Supervision
Our experiments are also carried out under professional guidance and supervision——our Principal Investigator, Professor Xingyin Liu, an expert in the department of pathogenic microbiology at Nanjing Medical University, She participated and guided our experiments from design to implementation stage, and join our weekly team meeting to follow up on the experiments.We have fully considered the possible risks of the experiments, and all agreed on Professor Liu’s suggestions on corresponding solutions. During the experiment, we can go to our laboratory technician Xiaoting Luan and the 2022 NJMU-China team member Yiran Wang for help. Xiaoting Luan and Yiran Wang are lab members of Xingyin Liu Lab and worked as advisors for our project this year.
In addition, our school, Nanjing Medical University, has a consummate laboratory management system , and we can always turn to the school for help if the risk presented turns out to be beyond our supervisors
Safety Training
Each of our wet lab members has passed the school's Experiment Safety Assessment and got certification before the iGEM competition.
Figure 1 Laboratory safety certification
Besides, we all have laboratory experiences and well familiar with the laboratory safety codes.Furthermore, each time before we carried out an experiment,we would received instruction and training of our PI.In order to use the lab during the summer break (July and August), we filled out the application form and got the consent of the college's laboratory management department.
Rules and Regulations
During the experiment, we have been following the rules below:
| Always wear personal protective equipment (PPE) and use appropriate gloves, glasses, and lab coat for each experiment |
| Use laminar flow cabinet to avoid microorganism contamination |
| Disinfect the surface with 70% ethanol before and after using the working table No drinks or food are allowed in the laboratory |
| Implement experimental garbage classification :different bins were used for different kind of waste |
| Label and store flammable and dangerous materials in designated areas |
| The use of an autoclave should be supervised by at least one graduate student Drug-resistant strains are carefully managed to prevent their release into the environment |
Human Practices Safety
Educational security
On the afternoon of April 28, NJMU-China went to Nanjing Fuzimiao Primary School to teach the children about synthetic biology and teach them to make leaf specimens. As scissors are needed for making specimens, in order to prevent the children from injuring themselves by mistake, we specially purchased round-tipped scissors and accompanied them throughout the whole process to ensure the safety of the children.
On the morning of June 10th, NJMU-China organized the high school students from the High School Affiliated To Nanjing Normal University to visit the State Key Laboratory of Reproductive Medicine of Nanjing Medical University. Due to the large number of people visiting the labs, the lab manager personally led the students on the tour, and members of the NJMU-China HP group and the team's instructor accompanied them throughout the tour to ensure the personal safety of the students and the safety of the labs. After the visit, we carried out the manual work of synthetic biology models in the conference room. We bought non-toxic and environmentally friendly ultra-light clay made of flour, water and vegetable pigment in advance. It has passed the China Compulsory Product Certification and will not cause harm to human body and the environment. The knives used to cut the clay are made of plastic, so there is no risk of scratches. During the entire process, each HP team member accompanies each student to make the clay safely.
Figure 2 is the picture provided in the details of the business, meaning that the scissors are round scissors, they won’t hurt hands and can prevent poking, they are very safe.
Figure 3 shows that the material and design of the scissors have passed the Tmall inspection, 3C, and EU triple certification, and met the standards for children's use.
Ethics
(1)Safety
Experiments in synthetic biology may involve gene editing of organisms or the introduction of synthetic gene fragments, so the safety of experiments and applications needs to be ensured. Ethical assessments should consider potential risks and hazards and ensure that appropriate safety measures are in place to prevent accidents or accidents from occurring.
(2)Social impact
The development of synthetic biology may bring about a wide range of social impacts, including economic, environmental and social changes. Ethical assessments should consider these impacts and try to anticipate and manage possible positive and negative consequences.
(3)Biodiversity and environmental protection
Applications of synthetic biology may have impacts on biodiversity and ecosystems. Ethical assessments should take into account the protection of natural ecosystems and the avoidance of irreversible damage to biodiversity.
(4)Fairness and justice
The development and application of synthetic biology technologies should be carried out in a fair and just manner and should not exacerbate social inequalities or create unequal distribution. Ethical evaluations should consider the potential impact of the research and application and advance the goals of sustainable development and social justice.
(5)Participation and informed consent
When conducting synthetic biology research or applications, it should be ensured that relevant participants and stakeholders are fully informed and consciously involved in the discussion and decision-making process. Ethical assessments should encourage transparency, informed consent, and public participation.
For more professional guidance, we spoke to Yong Li, Professor of Medical Ethics at Nanjing Medical University. Regarding whether the project is in line with current ethics in China, the relationship between synthetic biology and ethics, and his views on the current ethical problems in bioengineering and solutions, Professor Yong Li emphasised that synthetic biology research must be carried out under supervision and within a safe and controllable scope. When choosing topics, the risks and benefits should be measured to ensure that the project has scientific and social value. If our project is to be applied in the clinic or used for drug development in the future, Professor Yong Li stressed that we must have a complete ethical review and supervision system, and pay attention to the selection of drug materials for vulnerable groups such as children and pregnant women.
Laws and regulations
We fully respect the right of all interviewees to know and ensure the security of their personal information. According to the Personal Information Protection Law of the People's Republic of China, the processing of personal information should have a clear and reasonable purpose and should be directly related to the purpose of processing, in a manner that will have the least impact on the rights and interests of individuals. The processing of personal information shall ensure the quality of personal information and avoid the adverse impact of inaccurate and incomplete personal information on personal rights and interests. 1 According to the General Data Protection Regulation, the consent of the data subject means that the data subject freely gives specific, informed and explicit instructions by its will. 2 Therefore, we strictly follow the above requirements when completing the survey:
(1)First, no matter what kind of survey (online/offline interview, questionnaire survey, telephone interview, etc.), we will ask each respondent to sign informed consent form [ More details are available in Appendix I ] before the survey starts, so that they can understand the purpose, content and possible use of the information. They have the right to refuse any survey they do not want to accept, to refuse to answer any questions they do not want to answer during the interview, and to stop providing personal information at any time during the whole survey cycle.
(2)Secondly, the information we collect will only be used in the practical activities directly related to it, and will not be shared or used for any other purpose, not even in other activities of this project. For example, when we interview the families of children with autism, the content of the conversation between the two parties will only be used for our research into the lives and family situations of children with autism.
(3)Thirdly, during the interview and survey, some interviewees asked not to show their faces or appear in the camera. We met their requests by recording or mosaics, and sent the processed information to them for review. After receiving their consent, the information was finally presented on the website.
(4)Fourthly, we pay special attention to the protection of the privacy and security of special groups. When accompanying children with autism, we try to minimize the number of photos taken from behind or from the side, so as to avoid negative impact on them by taking pictures of their faces.
Figure 4 is a catalogue of the Personal Information Protection Law of the People's Republic of China, which refers to the provisions, rights, obligations, etc., of the processing of personal information. The figure is obtained from the China People's Net.
