Safety
A. Safety Concerns Involved in This Project
1. Safety concerns regarding the experimental host organisms
Escherichia coli (E. coli): E. coli is a minor component of the human gut microbiota and has the potential to be pathogenic. It can cause diseases when a person’s immune system is weakened or when the gut bacteria migrate to other parts of the body. Therefore, when using E. coli as a host organism for cultivating gene expression vectors, we must take this into account.
Bombyx mori (Silkworm): The silkworm is an insect from the order Lepidoptera, family Bombycidae, and genus Bombyx. Since it is not on iGEM's whitelist, we must fill out the check-in form accordingly. Although using silkworms as host organisms in transgenic experiments poses no significant risks to human health or the environment, we must also consider whether the experiment raises potential ethical concerns, the harm it may cause to the silkworm organisms, and whether alternative methods are available.
2. Laboratory level and corresponding safety measures
Our experiments were conducted in a specialized laboratory at Southwest University that focuses on silkworm research. This is a Level 2 laboratory (Moderate Containment). For high school students, a Level 2 lab environment presents increased complexities of equipment, environmental, and risk factors compared to the more common Level 1 labs in high school settings. This is a factor we must take into consideration before commencing any experimental activities.
3. Disposal of Experimental Outputs
In the final step of our experiment, we mix the transgenic silk powder with standard-sized PET plastic products to observe and measure its degradation effects. After the experiment, how should these transgenic silk powders and plastic products be handled? If the transgenic silkworms are released into the environment, could they pose potential risks to the environment? These are considerations we need to take into account as well.
4. Safety Concerns in Human Practice
Although our project does not primarily focus on social surveys, we are considering conducting one to better understand public attitudes and knowledge about PET plastics and their pollution within the local community. In doing so, we must take into account relevant laws, regulations, and institutional policies that may govern or structure this methodology. Additionally, careful consideration of the ethical implications of our survey is essential to ensure it respects participants' rights and maintains the integrity of our research.
B. How We Addressed Those Safety Concerns
1. Safety concerns regarding the experimental host organisms
Escherichia coli (E. coli): We used the Trans110 chemically competent strain, a commonly modified strain in genetic engineering experiments that poses very low risk. With proper experimental procedures in place, we can minimize the potential pathogenic risks.
Bombyx mori (Silkworm): Initially, we were unaware that using silkworms as a host for experiments required the submission of a check-in form. After being reminded by the iGEM officials, we submitted the form, highlighting an area for improvement in our future procedures and awareness. Given that the long-term goal of our research is to produce transgenic silk products capable of degrading PET plastics in the environment, as well as their overall advantages as host organisms, the use of silkworms in this project is unavoidable. We consulted laboratory experts, conducted background research, and concluded that silkworm embryo injection is a well-established technique. Additionally, PETase and MHETase are not toxic to silkworms, and their successful expression does not interfere with the normal physiological functions of the silkworm or produce harmful metabolic byproducts.
2. Laboratory level and corresponding safety measures
All our lab activities were under the guidance and supervision of the lab experts or qualified researchers. All personnel involved in lab activities wore protective clothing and other necessary personal protective equipment throughout the entire process. Before starting the experiment, they received a series of safety trainings, which included but were not limited to the following:
Safety Management in a Conventional Molecular Biology Laboratory;
Regulations on The Biosafety Management of Genetically Modified Organisms in Agriculture;
Southwest University Laboratory Safety Student Manual (including lab safety regulations, principles for handling laboratory accidents, safety in handling chemicals, toxic chemicals, and precursor chemicals, safe disposal of toxic and hazardous chemical waste, special equipment safety, gas cylinder safety, laboratory waste disposal, electrical safety, fire protection measures, and more).
The laboratory is equipped with sufficient safety measures, including sterilization and disinfection devices, biosafety equipment, waste management systems, chemical fume hoods, emergency procedures, and other necessary facilities and equipment.
3. Disposal of Experimental Outputs
The laboratory is equipped with a waste management system, and all experimental outputs have been properly disposed of. There were no issues of improper disposal or leakage of materials. Additionally, the survival capability of the experimental silkworms in the wild is very low, so even if they were accidentally to be released into the environment, the impact would be negligible.
4. Potential Safety Issues in Human Practice
Before conducting the survey, we referred to relevant laws and regulations of the People's Republic of China and Shanghai Municipality. We also consulted with the management of Shanghai Shangde Experimental School and confirmed that there were no regulations restricting us from carrying out this social survey. Nevertheless, we still need to consider the ethical legitimacy of the survey as well as potential risks regarding data security and data management.
First, our survey was conducted entirely anonymously. We collected respondents’ gender identity, age range, and education level to perform potential cross-tabulation of their answers. However, none of this information can be linked to individual respondents, nor will it be used in isolation for the research or any relevant release.
Additionally, we placed necessary restrictions on the participation of minors. All minors participating in the survey had to obtain consent from their guardians, with a dedicated checkbox included on the questionnaire for this purpose.
Furthermore, we ensured the maximum confidentiality of the collected data within our capabilities. Only a limited number of members from the Human Practice group had access to the survey data backend. The online questionnaire would be closed after a certain period, and the data was archived by designated personnel.
