Team:HZAU-China/Safety

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Overview

Safety has been attached great importance in every year’s project and experiment for HZAU-China, especially in 2021, still under the picture of pandemic. We not only abide by the safety rules and policies provided by the iGEM authority strictly, but also take every possible safety problem of our project into our consideration. It’s necessary to think about each aspect and give corresponding solutions as pre-arranged planning to avoid uncertainty, because our product will be implied into the actual working condition. Furthermore, since we are still under the condition of implementing the regular epidemic prevention, more attention was paid to reduce the human contacts and keeping the laboratory hygienic.

Lab Safety

Our lab locates in the main hall of College of Life Science and Technology, Huazhong Agricultural University. The main hall has an access control system, which can identify the student’s ID and only allow authorized students to enter (Figure 1). The teacher taking the charge of safety management of our college had been providing safety training for each team members and have routinly safety inspection for our lab.

Figure 1. Our access control system of entrance guard.

Safety Education

Professor Yingchun Qi is responsible for the safety and security of our lab. She has been working as the safety and security office in HuaZhong Agriculture University for decades, and she also gives lectures about biological experiments in university.

Safety of experience

Our laboratory is equipped with the fume hood, medicine cabinet (Figure 2). In addition, all the biological waste generated during the experiments is managed by trained stuff. So all of this measures can ensure that we can do experiment in a safe environment.

Figure 2. The safety of our laboratory. A. The overview of our laboratory. B. Fume hood. C. Trash bin for biological waste. D. Medicine cabinet.

All researchers wear gloves, masks, and lab coats (Figure 3 ). When handling volatile or irritating items, the experimenter is required to wear goggles to ensure safety. For using the instrument, the researchers strictly followed the instruction protocol to operate.

In our project, we need to verity the function of antimicrobial peptide (AMP) such as LL-37, which can neutralize lipopolysaccharide. So we used the Salmonella typhimurium SL1344 from Jin He’s lab in Huazhong Agricultural University, which has used in iGEM 2018 and sifA gene located in Salmonella pathogenicity island was knocked out (https://2018.igem.org/Team:HZAU-China/Safety). When operating the Salmonella typhimurium SL1344, the researcher was required to wear a mask, goggles and laboratory clothes, and used 75% alcohol to disinfect before and after the operation. Sterilize the operating platform with ultraviolet light after operation. Moreover, we had submitted the Check-in-form and got a permission. None of this would violate Chinese biosecurity policy.

Figure 3. A researcher operated with bacteria.

Safety of COVID- 19

Our project does not relate to COVID-19, and we did not use the RNA or protein of SARS-CoV-2. However, because of the COVID-19 pandemic, our team is taking a more cautious approach to offline human contact. All offline activities we hold were under the premise of confirming the health of both parties, and with the consent of the school and participants. We followed the epidemic prevention policy of the Hongshan district government, and all the activities are carried out with approval. During daily lab work and meetings, team members wear mask to minimize the risk. In addition, the laboratory is equipped with disinfection items, and the laboratory is cleaned and disinfected every day (Figure 4).

Figure 4. The disinfection items of our laboratory.

Safety of project

Safety is always the most important at every time. We not only about the safety of laboratory, but also focused on the project. We submitted the safety form, which including the detailed information about our project. the E.coli as our chassis organisms, which only make people diarrhea. The E.coli DH5α, BL21(DE3) were used to implement the experience of gene clone and expression. However, the above strains are often used in the laboratory, not for daily product. On the other hand, based on the result of questionnaire, we perceived that the safety of engineered bacteria is concerned. More than 50% of people worry about its safety. The safety of the project is ensured through both biological and physical methods. At the biological level, we design two suicide systems to ensure their safety, while for the physical safety, we use sodium alginate to embed our engineered bacteria. More details are shown below.

Safety in project design

At first, the chassis of our project is a widely recognized probiotic, E.coli Nissle 1917, which pose no harm to human health or environmental. Then, considering the implementation of our bacteria. We designed two different temperature-based suicide schemes. In the first scheme, the expression of HepT would be inhibited by heat-inhibition RNA thermometer in dogs’ guts, whereas it could be constantly expressed in vitro and make the bacteria commit suicide. In another scheme, we imported the HepT/MntA toxin-antitoxin system and kept HepT continually being generated. Instead of controlling HepT directly, we applied heat-activation RNA thermometer to activate the expression of MntA and neutralize HepT in intestine. Both schemes could perform the same expected suicidal function even if we employed diverse RNA thermometers with opposite effects. Furthermore, the specific suicidal scheme could be adjusted properly by changing the selection of RNA thermometers to tackle different room temperatures in different regions when it comes to practical application.

Safety of implementation

The microenvironment within the gel beads can provide protection and nutrition for micro. So that using gel beads has become very attractive for applications of our modified bacteria. We choose to embed engineered bacteria in sodium alginate, which can act as a physics barrier to avoid the bacteria escaping to the outer environment. Meanwhile, the bacteria have certain biological activity in the gel beads. Videlicet, we prevent bacteria from directly entering the environment without losing their biological functions.

In Compliance With iGEM's Safety Policies

“Do not Release Policy” is the core rule of our laboratory. First, we did not allow genetically modified organisms (GMO) to escape from our laboratory. At the same time, we have not released any GMOs into the environment. Secondly, there was no gene drive technology in our project. Furthermore, we did not implementation any animal experiments or human experiments. Finally, under the guidance of “Antimicrobial resistance policy”, we have not use any part containing sequences connected resistance to antimicrobials on WHO's list.