Our project will develop a c-di-GMP biosensor that can be widely used in different bacteria to quickly and real-time detect the concentration of c-di-GMP in living cell. To ensure the safety of the entire team during the iGEM season and to prevent accidents, we always follow the safety guidelines provided by the iGEM Committee to create a safe working environment. At the same time, in the era of COVID-19 pandemic, we have always adhered to the epidemic prevention rules formulated by the government, and actively and orderly promoted a series of experiments and activities under this benchmark. Next we will explain all the safety measures taken by our team to ensure the security of all involved parts including but not limited to classification of the microorganism, lab safety, operational safety, etc.

The two strains used in the project were Escherichia coli BL21 and Shewanella oneidensis MR-1. We chose them because they are model organisms for biofilm research and show low risk to human safety and the environment. In fact, according to the classification criteria provided by iGEM HQ, microbial risk levels are divided into four categories

Risk Group 1 organisms do not cause disease in healthy adults, and the risk to humans and the environment is negligible.

Risk Group 2 dangerous organisms can cause disease in humans, but the disease is treatable or preventable and poses low risks to humans and the environment.

Risk Group 3 organisms cause severe disease in humans. Treatments and vaccines for these diseases may exist and represent moderate risks to humans and the environment.

Risk Group 4 corresponds to operations that use transgenic organism from Group 4 and pose a high risk to human health and the environment.

E. coli BL21 and S. oneidensis MR-1 are classified as safety Group 1 by the American Type Culture Set (ATCS) and therefore require only one safety level 1 laboratory for handling. They belong to the White List set by iGEM for security.

Since the bacteria and the chemicals used in our project are non-pathogenic and non-toxic, the experiments can be conducted in Biosafety Level 1 (BSL-1) labs. Although we work in BSL-1 labs, basic safety procedures should be followed. For example, students working in the lab are required to wear long pants, closed shoes, lab coats and gloves. If necessary, wearing protective goggles is also required. Moreover, sterile operation is conducted in Biosafety Cabinet. The experimental area is completely separated from the office area.

Some common safety facilities and emergency devices

The bottles/tubes and our biological samples are properly labeled and placed in a certain order

Before leaving the lab and after handling biohazardous materials, hands must be thoroughly cleaned

The waste generated is classified and processed: the black garbage can mainly contains unpolluted garbage (e.g., sealing film), and the yellow garbage bucket contains biochemical drugs and other wastes such as culture medium, and test tubes.

The laboratory is equipped with sufficient personal protective equipment and has a counter for storage.

Biosynthetic research is inherently risky due to its nature. Under various security concepts of both laboratory work and the COVID-19 pandemic, a safe working environment was created for the safety of the whole team and the progress of the project.

Under the laboratory safety training, the operation safety was guaranteed, and the engineered bacteria containing antibiotic genes did not leak.

Before the practical project starts, team members had three months of laboratory safety training. As part of the laboratory safety concept by the CUG Lab, safety instructions were provided before the practical project starts and were always obeyed. Every member was equipped with protective equipment consisting of a lab coat, mask and disposable gloves.

The safety guidelines provided by the iGEM committee has also been adhered to at all times. All risks of wet lab procedures were assessed prior to practical work. For instance, the work with potentially hazardous chemicals, genetically modified organisms and equipment, such as disposable pipette tips and gloves. The above guaranteed that our engineering bacteria containing bio-bricks would not leak.

All parts utilized or designed in this project are on the iGEM whitelist or the necessary applications submitted. To prevent the spread of resistance to the antibiotics used, these antibiotics are used responsibly as a laboratory basis. In addition, the proteins encoded by our constructions, such as Green fluorescent protein (GFP) and transcriptional factor FleQ, are believed to pose no risk to humans and the environment. This project is in line with all safety requirements provided by the iGEM HQ.

Typical model organisms used in the project, such as E. coli and S. oneidensis, are not pathogens. Moreover, All experiments were entirely carried out in biosafety level 1 labs.

Collectively, the microorganisms used in the project are non-pathogenic, and the proteins expressed by our biobricks are not toxic. Therefore, the safety of this project is guaranteed.

Since the epidemic is still serious in this year,how to properly respond to the crisis and protect our own safety is not only a challenge to the world, but also a challenge to the preparation and experimental operation of the 2022 iGEM competition. To ensure the safety of our team in responding to COVID-19 pandemic, we always adhere to the COVID-19 rules setting by the government. This includes setting up team meetings online, wearing masks in the lab and during meetings, keeping social distancing, regular nucleic acid test, and following all safety guidelines in the lab space. Meanwhile, we closely monitored the vaccination situation and policy changes before we decided to organize offline team building activities, all of which were based on local rules for epidemic prevention requirements. Also, for those who are not well enough to attend the meeting, we provide the option to attend the meeting online to further reduce the risk of contracting other infectious diseases, such as colds. In addition, we limit the risk of contamination by regularly using water-alcohol gels on our hands and equipment. Fortunately, we didn't get any contamination