Since our project with its medical applicability can have a great impact on society, it was very important for us to have the consent of society regarding our methods. As we reached out to local groups, we realized that genetic engineering is still a controversial topic, surrounded with a lot of fear. To confirm this assumption, we conducted a survey addressing multiple questions around the topic and could conclude that with a lack of understanding comes a rejection towards genetic engineering. We took it as our responsibility to resolve as many uncertainties as possible that people from different communities have. The first place we visited were schools, where we could talk to the people of the next generation, because we believe this will have the greatest impact on the general understanding and attitude towards genetic engineering.
With our project presentations at different educational events, we tried to get many opinions on our project to include these into further development of our project.
The discussions and dialogues with the people after the presentation gave as an insight into opinions of people that are not that familiar with genetic engineering or scientific research and let us realize that proper science communication is important to build trust in new methods.
Apart from the education aspect we also got inspiration on ethic problems of our project and gave us the idea of trying another culture medium for Leishmania instead of Fetal Bovine Serum and through this minimize animal suffering as well as the impact on the environment.
Since our project aims to develop proteins, that can be used for research, therapeutics or vaccines, people were able to imagine the positive impact that our project could have towards medical research and drug development. In addition, people were impressed by the high security standards that we and our university employ and did not see any outgoing risks from our project for people or the environment. Scientists showed great interest in our project being the most directly benefiting group in terms of shortening their workflow in the lab and thus focusing more intensively on their research.
By working closely with a research group that works with Leishmania, we were able to adapt our project design directly to their needs. We could therefore directly implement their experience and insights into the development of our parts library. When contacting other researchers that work with Leishmania, we were glad to see that they show a lot of interest in our project and are willing to give it a try in their own research group.
With our communication and Human Practice work we were able to broaden our minds and advance the development of our project.
With our survey, we wanted to explore the opinions of people around the world on genetic engineering, as we felt it varies greatly from country to country. We couldn't think of a better way to reach as many people as possible than to ask the iGEM community for help. So we created a call for collaboration and asked all other teams to spread our survey in their country, at their university, among their friends and family. Unfortunately, we didn't get enough samples outside of Germany, so we couldn't draw any conclusions about how different nations compare, but we still learned a lot about German views on genetic engineering.
We had 219 participants in our survey.
Of the 219 participants, 109 were female and 110 were male.
With the age group, it was clear to us that we would be reaching mostly people our age, since the main platform for distributing the survey was online.
As expected, mainly people between 18 and 27 years old participated. More than half of our participants were in this age group, so our results may not be representative of all age groups.
Because the survey was spread across some iGEM teams and across our university, it was mainly individuals with an academic degree who completed the survey.
Now we know who participated, so let's take a look at the responses.
With this question, we wanted to find out how people rated their own knowledge about genetic engineering. Of course, this may not reflect their actual knowledge because some people tend to overestimate or underestimate themselves.
We wanted to find out what the attitude of our participants is towards genetic engineering, because we had the impression that the attitude is often rather negative, because people are biased by the critical reporting in the media on this topic.
Even though we thought people were more negative about genetic engineering, we know that when it comes to medical relevance, opinions can be somewhat different.
We wondered if the biased attitude toward genetic engineering might result from a lack of education about the topic, and we wanted to find out how people think about education these days.
The results of the first question were as expected, with neither very high nor very low knowledge (MV: 2.79, SD: 1.239). To our surprise, the average attitude was slightly positive (MV: 3.63, SD:1.172). As expected, people are more open to genetic engineering in medical contexts, which can be seen from the fact that the answers to question 11 are more positive. (MV:3.85, SD:0.982) On average, education about genetic engineering is considered rather insufficient (MV:2.04, SD:0.918).
As mentioned earlier, we expected that attitudes toward genetic engineering would differ between the different application areas.
As can be seen, genetic engineering meets with significantly more opposition in agriculture than in medicine or industry.
Since genetic engineering in agriculture has a direct impact on people's daily lives, this area of application tends to be more important to people. One situation where they would likely align their actions with their values would be when shopping.
32% of respondents said they look for the "NO GMOs" label when shopping, which is consistent with the 68 people who think GMOs don't make sense in agriculture. The 151 other respondents who answered "YES" to question 3 are similar to the 68% who don't care about this label.
The next questions were about the legal situation and how people want to see it.
Although 31 % of respondents are not positive about genetic engineering in agriculture, only 29% want it banned. Surprisingly, more people want to ban the import of genetically modified crops than want to ban their cultivation. Also surprising was that 87% of respondents want GMO foods to be labeled, although only 68% pay attention to the label when shopping.
27% of respondents believe that GM foods are harmful to their health. In this question, we also asked about the ways in which they think the food can harm them. Most people did not give an answer to this question.
Of those who did answer, six people did not think of the direct effects on their bodies, but rather environmental effects that also affect their health in more indirect ways. Besides these, allergies, cancer or toxic effects on human metabolism were also mentioned.
Further on, the topic was the use of genetic engineering in medicine.
As seen before, the use of genetic engineering in medicine is more accepted, which may be because people see a direct positive effect on health, as can be seen in the graph below (MD: 3.85, SD:0.98).
We also wanted to know where exactly people can see the effects of genetically engineered medicines.
As you can see, people see a lot of potential in genetically engineered drugs, especially in terms of cancer treatment, but also in many other diseases. These include inherited diseases, neurodegenerative diseases, autoimmune diseases, and vaccine development.
In addition to the great potential seen in the application of genetic engineering in medicine, some risks are also seen.
We also offered an open area where respondents could list other risks, but most fear misuse of these medicines as well as lack of experience with their use. As you can see, many of the risks people see with genetically modified drugs are risks that could apply to any other drug.
We now know how people on average feel about these issues, but the interesting question for us was why people often have negative attitudes toward genetic engineering, and although the survey showed that this was not the case for our participants, we were still able to draw some conclusions.
First, we wanted to know if age made a difference in knowledge, attitude, or view about genetic engineering education.
The correlation test showed a significant negative correlation between age and estimated own knowledge about genetic engineering (r= -0.202, p=0.003), which is a weak effect according to Cohen (1992). That is, the lower the age, the more individuals believe they know about the topic. It is the same case for attitudes toward genetic engineering (r=-0.335, p=0.0), which resembles a strong effect.
It can be concluded that younger people tend to know more about genetic engineering and have more positive attitudes toward it. We also ran a correlation test with age and education scores, but there is no significant difference.
Does this also mean that more knowledge correlates with a better opinion of it? Correlation tests showed YES!
There appears to be a significant correlation between more knowledge and a more positive attitude towards genetic engineering (r=0.478, p=0.0). This is a strong effect according to Cohen (1992).
There is also a correlation between degree and knowledge. According to Cohen, this is a medium effect size (r=0.238, p=0.0). The degree also shows a weak correlation with attitude toward genetic engineering (r=0.146, p=0.0). This means that individuals with a higher degree rate their own knowledge higher and have a more positive attitude towards genetic engineering. Individuals with higher degrees also tend to rate genetic engineering education higher. According to Cohen (1992), there is a weak correlation (r=0.188, p=0.0).
We also wanted to test whether there was a difference between the genders. The Mann-Whitney U test showed that there is no significant difference between men and women in terms of knowledge, but men seem to have a more positive opinion of genetic engineering. According to Cohen (1992) it is a small effect d=0.237 (U=5025.5, Z=-2.319 p=0.032).
People who believe they have a higher level of knowledge tend to have better attitudes towards genetic engineering. From this, we can conclude that many of the prejudices or fears that exist about genetic engineering can be reduced through better education. The results also show that younger generations seem to be better educated (! Keep in mind that we did not have a large sample in the older age groups) than the elderly, which could be the result of better education nowadays, but this seems somewhat unlikely since there is no significant difference in education scores between the age groups. The other reason could be that younger people have the information more readily available in this age of digitalization and are more accessible to obtain information.
All correlations were tested using Pearson correlation. Effect sizes were calculated according to Cohen (1992). The results may not be representative because we had a small sample and reached many individuals in our own peer group. Because this includes many biologists and other iGEMers, responses may be biased.
Integrated Human Practices
While working on our project, we faced some difficulties and turned to experts from different companies to help us solve these problems. Besides helping us with our experimental problems, we also talked about the implementation of our project and which criteria our project must fulfil in order to be applied in the real world.
Dr. Andreas Licht - Jena Bioscience
Our target vector is based on the pLEXSY_blecherry3 plasmid provided by Jena Bioscience. Dr. Andreas Licht is the expert in charge of this system at his company. When we tried to domesticate the plasmid, we had problems with GC-rich sequences that we could not replicate by PCR, so we contacted him to meet us. But aside from that, he gave us a lot of advice on protein expression and transfection of Leishmania that we had not yet considered. His experience working with Leishmania was very valuable to us through the tips and tricks he showed us and really helped us move our project forward.
With the help of all his expertise, we were finally able to successfully transfect our leishmania and detect the expression of mCherry, which is an indicator of the presence of our plasmid in the colonies. However, successful transfection alone is not proof of concept for our project and therefore our goal was to purify our example protein the receptor binding domaine of the Sars-CoV-2.
Since all of our constructs have purification tags, we thought that purification should be easy, which ended up not being the case. After much trial and error, we contacted Dr. Licht again and he gave us advice on how to properly culture Leishmania to achieve high protein expression. We learned that when diluting cultures, we had better dilute more often, but with lower factors. In addition to practical tips, we also talked about the long-term implementation of our project.
We considered who our target audience would be. Our first thought was large-scale protein production of human-like glycosylated proteins, but Dr. Licht's experience showed that protein expression levels in Leishmania tarentolae are usually too low, so our target audience would be scientists rather than industrial companies.
Our modular system allows scientists to easily incorporate their desired protein and provides them with many ways to detect and purify it, making the system very suitable for scientific work.
Dr. Licht also gave us some insight into problems he is experiencing or has heard about with the LEXSinduce3 Expression system. One problem is that some genes just don't seem to integrate into the vector. It seems that our modular system may be the perfect solution to this problem, as modular cloning reactions lead to a higher number of successful transformants.
Dr. Christian Janzen
Our second PI Marcel Deponte put us in touch with one of his former colleagues who was also working with Leishmania. We got in touch with him and learned that he had unfortunately stopped working with Leishmania because he was having problems. We set up a meeting to find out what had gone wrong.
Christian Janzen and his graduate student Elisa Theres Rauh explained to us that their goal was to express Trypanosoma proteins in Leishmania tarentolae because they are very similar organisms and typical expression hosts like E. coli have difficulty mimicking the complex proteins of Trypanosoma.
We had to deal with one of its problems, which is that the genome of Leishmania is very unstable. They were able to detect the proteins they were interested in via Western blots, but only in new cultures. After a few weeks, the same cultures showed no expression, leading to the assumption that Leishmania is very sensitive and deletes the genes it doesn't need. From this, we learned why perhaps our very first purifications didn't work, because the cultures we used for them were quite old.
In addition, Christian Janzen and his research group also had problems with cloning. Some of the gene sequences for the proteins they wanted to express simply could not be cloned into the vector. The group was working with the same expression vector from Jena Bioscience, so we agreed to share our domesticated vector with him so that he could try out whether this solved his problem.
In addition to talking about his problems, we also shared our own problems with purification with them. Since we had the problem of our GST tag being cleaved off in Leishmania, they gave us the advice to use a special blend of protease inhibitors, since the commercial blends often don't contain the inhibitors you need. We plan to incorporate this advice into our upcoming work on our project after the iGEM competition.
We greatly appreciate your support, thank you again to Dr. Christian Janzen and Elisa Theres Rauh.
Interview with Matthias Mieves
In September 2021, the Bundestag elections took place in Germany. As this election was very decisive for the future of Germany, the topic kept us all very busy. In the middle of the iGEM project, the question naturally arose: What are the parties' positions on genetic engineering?
To find out, we took a look at the election programs and analyzed the positions not only for us but also for a broad audience via social media.
This made us notice that the positions are almost consistently negative and that genetic engineering is often unfoundedly rejected, especially in the field of agriculture.
In order to investigate this problem from the ground up and get to the root of it, we decided to invite a local politician to talk to us. Matthias Mieves (SPD = socialdemocratic partie of Germany) agreed to talk to us and came to visit us in the lab. After a quick tour through the lab, we also interviewed him about his opinion about genetic engineering.
In general, we found out that he is very open minded towards genetic engineering and also sees the obligation for the government and politicians to support researchers financially but also with communication. Matthias Mieves thinks that one big problem about communication and media is that most of the articles about controversial topics in general and of course about genetic engineering only display the risks and fears that people have about it. Companies, media and politicians should also report about their successes in research or the big opportunities and possibilities that come with genetic engineering. One of those big opportunities and advantages of genetic engineering he sees is when it comes to the global food supply, where genetic engineering could be a big part of the solution because with genetic enigeered plants, we have more possibilities with cultivating plants that are more resistant to drought or heat.
To educate people more about genetic engineering and get rid of the fears people have, Matthias Mieves wants to work with clear communication and transparency and not force anything onto people. For transparency reasons also the “NO GMOs” label is a good way to keep people informed and give them the possibility to make their own choices, but at the same time the government can still support further research and innovations. On the other hand, he does not support a “GMO” label because he does not see where this would help anyone and where to draw the line since animals nowadays are also fed with GMO modified foods and are also bred to be more productive, e.g. produce more milk or grow faster, which also is a form of genetic engineering. Also, a label like this would promote the stigmatization of GMO foods. From another perspective, this does also make no sense, since many foods and especially highly processed foods contain a lot of additives and a lot of sugar and fatty acids that are not healthy or even proven to be unhealthy, but are also not labelled. A more transparent system should rather inform consumers about the risks of the food, than state a fact like “contains GMO” which does not pose a threat to their health.
Apart from health issues, we also talked about the social aspects of genetic engineered plants. One big problem is that big companies produce the engineered crops and make a lot of money with it. Often the crop is sterile which means that farmers have to buy it year after year after year and go bankrupt with this, since the company that produced it, has a monopoly on it and can dictate the prize. Matthias also sees the need to prevent this and thought of handling it like we do with medicine and patent protection, where a company has a patent on their new product for 5-7 years to get their money back that they used for research and production, but after this the patent expires and every other company can use the technology. This way, the money keeps the companies motivated to do further research and find new products, but do not get a monopoly on anything.
Apart from his very open mindedness about the topic and his supportive attitude, Matthias also sees some risks regarding genetic engineering. One is the safety aspect e.g., regarding GMO foods that must be tested properly before they get out on the market, so consumers stay safe and do not take any harm. Second one is a risk that comes up with all new technologies or innovations: the risk of misuse. In our survey this was also the most concerning thing for all participants and is probably the reason for the very negative attitude of the german parties. Matthias does also see this risk but doesn’t want to let the fear of this dominate the debate about it and wants to find a way to let researchers do their work without being impacted by the rules around it, which he at the moment still sees in Germany.
Genetic engineering is not only used in agriculture but also in medicine, so we also asked for his opinion on this as well. As expected, he was also very open minded about this but also sees the need to be careful with this, especially when it comes to germline mutations, because any modification here becomes irreversible.
We hope that Matthias Mieves as a new deputy of the Bundestag will be able to spread his opinion throughout the SPD and get Germany to a more supportive position towards innovations and new technologies, especially genetic engineering. At this point we want to thank Matthias Mieves for finding time to have this really nice talk with us and wish him good luck with his time in the Bundestag.