Safety has always been the first priority of our team. Being transparent about possible risks and how we are managing them is a key requirement for responsible scientists or engineers. This year, Tsinghua iGEM team comprehensively analyzed the possible safety risks encountered in the project and daily experiments, both inside and outside the lab, to demonstrate our understanding of "responsible design".

Biosafety Laws in China

The Biosafety Law of the People's Republic of China was adopted at the 22nd meeting of the standing Committee of the 13th National People's Congress on October 17, 2020. All of our work must obey the law.

Training

Each year's iGEM team members have been systematically educated on laboratory safety. In order to gain lab access, new members were required to attend an offline course and pass an online exam. The required courses included modules on basic laboratory practices, chemical use safety, biosafety education and GMO safety, etc.

In addition, our PI and Tsinghua iGEM 2021 team members conducted another detailed lab safety training for our iGEM lab after the iGEM members were confirmed for this year. The content included personal safety protection, waste disposal standards, biosafety precautions, laboratory electricity and fire safety, etc.

Lab Safety Rules

After each lab member has received basic lab skills training and familiarized with the lab, we further developed our "Lab Co-Building Rules"based on standard rules proposed by the previous iGEM team and our own revision. The Lab Co-Building Rules were the embodiment of every iGEM member's participation in the construction and development of the lab. These rules have been strictly enforced during the whole year of participation in iGEM.

1. Equipment use and consumables access: in this section we list the rules for the use of commonly-used instruments and laboratory consumables. In particular, we list the potential dangers and describe in detail how the risks should be minimized through laboratory practices.
2. Personal duty: We break down the daily tasks that need to be accomplished to maintain the cleanliness and safety of the lab, and assign the work and responsibilities to individuals. Such work division not only helps to improve the efficiency of the lab, but also helps everyone to understand the effort required to maintain a properly functioning lab, thus raising everyone's consciousness to maintain the order of the lab and respect the faculties who provided us with such a good laboratory environment.
3. Last person: Following the experience of iGEM team in previous years, we continue to revise the last person rule of the lab, which requires the last person leaving the lab every day to check the instruments that should be turned off in the lab to avoid unnecessary waste of electricity. Meanwhile, we pay close attention to the operation status of common instruments to minimize the safety risks in the lab.

The last person rule has been well implemented throughout the whole year. It not only records our hard work in the lab every day, but also allows us to clearly see who really does experiments until late.

The major safety concern of our project stems from the engineered bacteria and materials that we proposed to be administrated into living creature.

Microorganisms Review

Based on the White List and Risk Group classification of microorganisms permitted in iGEM, we selected E. coli strains that have been widely used and demonstrated to be safe to conduct our experiments. Strains involved in our experiments are listed in the following table.

Reagents Safety

In our project, in addition to the commonly-used IPTG and arabinose, the inducers involved in the functional verification of the parts include antibody and nisin, which are required for our self-designed two-component system of Pmr and Nisin. The antibody can be any antibody with an intact Fc fragment, which is essentially an immunoglobulin and has a low risk level during the experiment. As for nisin, often used as a antimicrobial peptide, we conducted thorough literature research about the potential safety concerns of nisin before conducting our experiments.

Nisin, an antimicrobial peptide produced by some species of Lactococcus lactis, has broad antibacterial spectrum against Gram-positive bacteria species. The use of nisin has not lead to any problem of widespread resistance. Toxicological tests in various countries have proved that YP-1000 Nisin is safe and non-toxic.

In addition, according to our design, our fusion protein NisA-affi is produced by fusing affibody with the C-terminal end of NisA. Our results demonstrated that the fusion protein would not change the original properties of NisA or generate any harmful substance. In conclusion, our design would not cause any safety concern.

Engineered Bacteria Safety

Since the engineered bacteria will be present inside our microfluidic chip, we need to consider the environmental risks of our strains when released to the environment to comply with the green development concept and requirement. To achieve this, we could design our strains to be auxotroph so that they are unable to live outside our device.^[1] In this way, even environmental release happens, the engineered strains will not cause much harm to the environment.

Hardware Safety

Material

We try to choose non-toxic, odorless and environmental-friendly materials when constructing our hardware device. PDMS (polydimethylsiloxane) is a thermoplastic synthetic resin, which is colorless, odorless, non-toxic, and translucent material with good heat and corrosion resistance. Therefore, PDMS can be an ideal material for our microfluidic chip base, ensuring a stable detection system without any potential leakge to the environment.

Disposal

After our microfluidic chips have been used, users are recommended to inactivate the engineered bacteria inside the chip by treating them at temperature above 65°C to avoid potential leakage. Previous iGEM teams have designed inactivation switches such as "self-heating package" (NEFU_China, 2021) to prevent potential biohazard contamination leakage to the environment. Such design could be introduced into our product as well.

Data Security

In the course of our Human Practice process, we designed a questionnaire on public perceptions of current sperm quality testing products. The questionnaire was designed, distributed, collected and analyzed in accordance with the principles of the General Data Protection Regulation (GDPR), which protects the privacy of participants.

Lawfulness, Fairness and Transparency:

Our data were processed lawfully, fairly and with the survey participants' consent. We clearly informed the participants of the subject matter and general content of our questionnaire before we administered it, and obtained their informed consent.

Purpose Limitation:

We designed the questionnaire in such a way that only information about the public's perception of the sperm quality testing device and the most basic identifying information about the participants (e.g., gender, age) would be collected. No information about the participants' personal health status would be included.

Data Minimization and Accuracy:

We designed the questionnaire to ensure that only information relevant to the topic was collected and that it was limited to this study. Our questionnaire was also designed to ensure that the information collected was as accurate as possible.

Storage Limitation:

We plan to delete the raw data of this questionnaire 6 months after the Jamboree, keeping only the statistical and analytical results without any particular personal data.

Integrity and Confidentiality:

We try to ensure the security of the information in the process of storage, such as storage in password-protected computers, handling and storage by dedicated personnel, avoiding the transmission of raw data through online social networking software, etc.

Accountability:

Our team is committed to take responsibility for this survey.

[1]Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., & Gennis, R. B. (2015). Escherichia coli auxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods in enzymology, 565, 45–66. https://doi.org/10.1016/bs.mie.2015.05.012