Q1：What values-environment, social, moral, scientific, or other-did you have in mind when designing your project？
Gene editing is an important tool for synthetic biology. In our project, we modified the signaling pathway in yeast and tried to use CRISPR/Cas9 to knock out the ste5 to construct mutant strain. We designed a questionnaire about CRISPR technology, hoping to get some valuable feedback from other iGEM teams. In addition, we performed campus interviews to investigate perception and attitudes of gene editing among students of different majors. Feedback from these target population helped us better evaluate the environmental, social, moral values when designing our project.
A1：A questionnaire about gene editing
The following pie chart shows some iGEM teams participating in the survey.
We did an investigation into gene editing, targeting at iGEMers. 57.89 percent of those iGEMers who participated the questionnaires have used CRISPR in gene editing, which is a splendid usage rate for the emerging technology. We also looked into why some iGEMers don't use CRISPR. There are a lot of iGEMers who don't know how to use CRISPR, and some labs don't use CRISPR because they don't have plasmids, so we will share CRISPR plasmids with them. For iGEMers, only 9.09 percent of those who have used CRISPR have used it as a molecular switch, so this is a novel and creative bright spot to look at. Besides, the safety issues of gene editing are also the focus of people's attention. Most iGEMers have considered the risks of gene editing on human health and environment. The ethical issues of gene editing influence their selection of experimental objects. The future of gene editing is highly uncertain. Therefore, the application and development of gene-editing technology must be taken seriously.
If inter-species cells fuse, it will be a good thing for human development to produce superior new species.
Nanjing high school student:
I think if it is a technological advancement, corresponding research can be conducted. I support the theory of progress of the times, but at the same time, there should be absolute rights to standardize the management of such scientific research activities to prevent illegal individuals or groups from using genes to perform actions that are harmful to humans. In addition, I personally think that there are no ethical issues at all. Thoughts need to follow the progress of the times. Staying in place will only be eliminated. Humans and new genetic organisms can coexist harmoniously after a period of running-in.
Electrical engineering junior:
It is understandable that genetic modification and cell fusion are purely to transform the development needs of human beings in the future. But people's pursuit of profit has raised questions. Using Taoist thinking, governing by doing nothing, letting nature take its course, and deliberately changing what nature gives, there may be devastating "disasters”. Everything has its opposites, there is good and bad, in summary, I am cautiously optimistic about this issue.
International trade junior:
From a commercial point of view, when genetic characteristics are well optimized, it will very likely to replace the value of the original crop in the market. Meanwhile, the birth of industries and related derivative industries is similar to the promotion of the industrial revolution to the development of productivity. The birth of good genetically modified products is bound to be revolutionary for the market. However, genetically modified products are very uncontrollable from a biological point of view, and the impact of new gene combinations on the gene sequences of all organisms is unpredictable. Therefore, when genetically modified technologies are put on the market, consumers may not be very willing to accept these products.
Pharmaceutical engineering junior:
The cloned sheep Dolly is a new species generated by cell fusion and incubation, so she only had six and a half years of life, finally died of pneumonia. Therefore, although the new species and new traits produced by cell fusion are beneficial to human development, it is necessary to improve the technology of cell fusion and subsequent culture methods.
Environmental engineering junior:
For genetic modification, I feel that it’s okay to eat genetically modified food as long as there is no problem with it. Other ethical issues should be considered by researchers. Genetically modified food can be eaten if it is harmless to human health. Nowadays, I feel that many people have a lot of prejudice and misunderstanding about genetic modification. I also hope that scientific researchers can clarify, answer everyone's doubts, and eliminate misunderstandings. In addition, if genetically modified animals or plants are cultivated on a large scale in the outside world, will they have some impact on native species? These are safety issues that need to be considered.
Public health junior:
Transgenic and cell fusion can meet some of the special needs of humans. In this way, it can indeed help human life to develop toward an ideal situation. But to a certain extent, this technology has caused the destiny and laws of life to be broken, and may even make humans lose the meaning of life and threaten human health. Therefore, in the process of supporting the further development of transgenic and cell fusion technologies, it is also necessary to grasp the scale and establish an effective monitoring and management mechanism to prevent the dangers brought by technology, so that it is the basis for ensuring the safety of human life and the meaning of life safety development.
For the college student community, we conducted a series of campus interview activities. College students of different majors have different attitudes towards genetic modification and cell fusion. Some college students believe that genetic modification will make human genes converge and trample on moral standards. Some college students believe that genetic modification will disrupt the laws of life, making humans lose the meaning of life, and even threatening the long-term development of human beings. While some college students hold the opinion that scientists should clarify the advantages of genetic modification as soon as possible and let humans abandon stale concepts and live in harmony with genetic modification. Because our team project involved gene editing and other operations, we also talked about the safety of this type of genetic modification in later science education. We took the safety issues into account and found that some college students have a clear understanding of genetic modification technology.
Q2: Which resources of communities did you consult to ensure those are appropriate values in the context of your project?
A2: We contacted the Center for Disease Control and Prevention of Wuxi Municipal Health Bureau to inquire about the causes and harms of the spread of resistance and antibiotic contamination, because yeast mating can cause unexpected gene spread.
The abuse of antibiotics has aggravated the contradiction between antibacterial drugs in a balanced state and bacterial resistance. Bacteria with drug resistance have also acquired the ability to resist different antibacterial drugs through continuous evolution and mutation. This ability is in contradiction. The struggle is constantly being strengthened, and bacteria gradually gain multi-drug resistance or even pan-drug resistance from single-drug resistance, eventually become drug-resistant super bacteria.
Bacteria mainly resist the action of antibacterial drugs in six ways. Produce hydrolase or modified enzyme to hydrolyze or change the structure of antibacterial drugs and inactivate them. Change the target position or number of antibacterial drugs to prevent them from combining with antibacterial drugs. Modify the permeability of the cell wall and prevent the antibacterial drugs from entering the bacteria. Through active efflux, the drugs are excluded from the bacteria. These bacteria secrete extracellular polysaccharide protein complexes to surround themselves to form a bacterial biofilm. Integrate the subsystem. One of these resistance mechanisms can make bacteria resistant, but they do not exist in isolation from each other. Two or more different resistance mechanisms interact together to determine the resistance of a bacterium to an antimicrobial drug level.
In the communication with the CDC, we learned that the abuse of antibiotics will aggravate the contradiction between the balanced antibacterial drugs and bacterial resistance. Many bacteria have evolved and mutated to form so called super bacteria. We have also learned about the ways in which some bacteria resist antibacterial drugs. For the project in our laboratory, we used kanamycin and ampicillin, and we realized the seriousness of antibiotic abuse. Therefore, in our experiment, we will pay attention to the classification and treatment of laboratory waste such as tablets to avoid harm to the environment. In the selection of plasmids, we will tend to choose plasmids with auxotrophic marker gene to reduce the use of antibiotics.
A2: Professor Xin Wang
Professor Xin Wang suggested that it is meaningful and feasible to use galactose instead of pheromone to induce yeast expression and reduce the cost of induction in our project.
Therefore, we continued constructing a signaling pathway mutant in the project and used galactose instead of pheromone to induce the expression of pheromone responsive genes.
A2: Professor Dong Liu
Professor Dong Liu suggested agreed that in some cases, the modification of promoters is better than the changing of copy numbers for gene expression regulation. We use promoters to regulate gene expression instead of using multiple copies for over-expression. Multiple copies are unstable, and the use of promoters to regulate and control the effect will be more significant. Professor Dong Liu also raised his own question consideration regarding the cost advantage of galactose-induced yeast expression: the replacement of pheromone by galactose would switch the carbon source, so there would be some influences on the growth of yeasts in cell growth.
We compared the regulatory advantages of various promoters and selected three promoters pprm1, pfus2, and pfig1 for use deep research in our team's project. Take Professor Liu’s suggestion into consideration, we will also plan to use light control or temperature control responsive promoters to solve the problem of cell growth.
A3: Professor Xiaojun Ji
Professor Xiaojun Ji proposed that comparing to changing the copy number of genes, promoter regulation is a more refined and stable regulation. He suggested that we learn more about promoter engineering and its practical applications.
A4: Professor Fengxue Xin
1. With respect to the practical applications of your system to biosensors, specific detection targets need to be considered. Antibiotic in the environment provides a possibility.
2. Environmental monitoring usually requires low detection limits. Since you are considering applying the pathway to environmental contaminant detection, it is worth considering how to improve the sensitivity of the detection system.