Human Practises for us means to show our project to the public, our excitement for it and to encourage as many people as possible to think about and participate in the discussion regarding synthetic biology and the various aspects linked to it. At the same time, it also means to get external input on how to further improve and adjust our project, solve problems and work together as a team effectively and having fun on the way. On the following page you will see what we have accomplished throughout the year. We hope you will enjoy reading!

This year we chose a project that contains a highly potent neurotoxin: the botulinum toxin - shortly botox. In order to fulfill the iGEM lab safety guidelines we had to work with a detoxified version of the protein allowing us to work in a S1 laboratory. We had to overcome a few obstacles before we were able to do so. For this purpose, we stayed in contact with experts of botulinum toxin throughout the year and accompanying to our project. We gained a lot of important knowledge and invaluable advice from the correspondence in this research field. It also offered us the opportunity to modify certain parts of lab work and the chance to restructure our project. Due to the advice of experts like Dr. Binz we learned a lot about how to handle toxic substances in the lab and came up with a safety concept. Through interviews with politicians, medical doctors and scientific researchers who gave us constructive feedback and some new insights we got to reevaluate the potential of our project, possible areas of use and smart additions to it.

Identifying the Risk Factors and -Level

After we found our project we knew there would be difficulties concerning safety and handling as botulinum toxin is one of the world’s most toxic substances, listed as a potential bioweapon. As we tried to discover how we could work with it nonetheless, we came across the possibility to insert specific mutations which render the protein atoxic. Originally, we planned to order our constructs at IDT. In Hannover, we met Dr. Binz, whose paper inspired us to do our project in the first way, and talked about Botox in general, its characteristics, how his group works with the toxic version and talked also about secure work. We also discussed the necessary mutations to detoxify the protein and asked whether he thinks they pose a valid method of doing so. He gave us invaluable tips on how to accomplish those changes of the DNA sequence. We also discussed our coupling plans for the additional proteins, which would normally be considered as S2, but our work was regarded as S1 as we only worked with DNA. After our botulinum toxin was certainly detoxified, working with the protein was also considered as S1.

Consequent upon our exchange with Dr. Binz, we had a useful talk with Dr. Kittel, the biosafety officer of our university. We discussed whether we were allowed to work on our project in an S1 environment. It turned out that working with DNA only is appropriate for S1 and that once we have proven via sequencing that the detoxification works well, we were also permitted to work with the whole proteins in S1 surroundings.

Following the allowance to work in S1 in our university, we talked with our PIs and presented our ideas and how we planned to handle our constructs. As our PIs and professors were concerned that the ordered botox with the mutation would not be sufficiently detoxified, they didn’t want us to work with the IDT parts. Luckily, we were able to obtain the wt plasmid from Dr. Binz. On DNA level, we cleaved the light chain (LC) from the heavy chain (HC) and our PIs stored the HC. We had only access to the in itself atoxic LC. Throughout our project, we always handled LC and HC separately and the wt plasmid was also stored by our professors. Our professors also contacted the regional council. We agreed that the plasmid was to be divided in HC and LC in an S2 area by an experienced professor. We were only given our Botox LC once it was safely divided. At this point, we were allowed to work with our Botox LC with the admission from our PIs, our university and the regional council.

The iGEM Safety Committee

As it is especially important for iGEM that the projects of their contestants are safe, we asked the committee whether we were allowed to continue with our project and gave them all our available information about bioweapons and detoxification among others. In their answer they agreed that once we were able to prove the detoxification successful, we could work with it. But our submitted parts will not be included in next years distribution kit, due to possible toxicity. We also had to fulfill the safety sheet and check in sheets (iGEMs safety forms) thoroughly. Additionally we talked with the safety officer at the european meetup in person. Through this procedure we wanted to completely insure that we could work with our project savely. Our next issue was that we were working in two different labs in two different institutes. We needed a security clearance and talked with Mrs Fuss who is responsible for the safety in the cell culture department because our assays needed to be executed with our cell culture and we therefore had to work with botox in the lab. According to her, we were only allowed to work with proteins in her lab, not with DNA. As we ensured the director of the interfaculty institute for biochemistry Prof. Dr. Jansen the safety of our project, an additional concern from another professor came up: we had to be careful of how our project would be presented in the media (concerning risks and possibilities).

University and Institutes

After ensuring our institute director that we thought about all concerned security measures, we had to inform him and all other working groups about the major processes of our project and we had to inform him whenever we started the next step.

The Teammembers

To carry out our project successfully, we had an introductory seminar about protein purification, held by Frank Essman. According to him, we always have to keep in mind that we are working with a highly toxic protein hence the necessity to apply the highest safety measures in laboratory work. Our team needed to work as careful with our botox as if it was still toxic. The next part was our contact with professor Stehle who helped in purificating our proteins. As the protein is potentially toxic, this was one of the most important aspects. We held a presentation in front of professor Georg Zocher, who coached our work, and the biosafety officer of our working group. Together we concluded that the knockout criterion had to be that we should be able to proof, by sequencing our protein construct, that it had the intended mutations and DNA sequence. And only after we had validated this through our sequencing results, we were allowed to transfer our construct in E-coli, express and purify our protein.

To sum up, the three most important points you have to keep in mind when working with toxic substances are the following: First, stay in contact with iGEM so you can consider their advice and guidelines. Second, clear everything with your university so that your work takes place under the right circumstances. And last but not least, make sure that all members of the team work with the right mindset and carry out their tasks carefully - with the potentially high risk in the back of their heads.

One of our main focuses for this years project was to increase our public outreach significantly compared to the previous years regarding our social media performance as well as the organisation and attendance of events on a local level, all over germany and also europe, and getting in touch with scientists and politicians.

Ract! Festival

We addressed people of all ages. To introduce kids to biology and raise their interest we build a parcour that abstractedly symbolised the shuttle mechanism of the botulinum toxin into neurons. It turned out to be a huge attraction. To reach out to adults we passed out flyers for our upcoming public debate and postcards of previous iGEM projects. Additionally, we were happy to answer questions and offer new insights. We were pleasantly surprised how many people sympathised with our believes and the potential of synthetic biology. Nonetheless, we had some negative encounters with opponents of genetic engineering. We hope that we at least encouraged them to reevaluate their opinion. All in all it was a great success.

Public Debate

Following to the Ract! festival we organised a public debate that took place in our university. This event was organised in collaboration with Streitkultur e.V., the debating club of our university. It was a public debate about gene editing on embryos which is a highly controversial topic in today’s society. We had two mixed teams, each with one iGEM member and one member of Streitkultur e.V. that represented the advocates and opponents of gene editing. After an introductional plea of both sides the audience had the opportunity to ask questions and take part in the debate. They were very inquisitive and we had a lively debate in a friendly and respectful atmosphere. It was a great opportunity for our team members to improve soft skills concerning confrontations with opposing opinions and how to handle them. The debate served the purpose of making the subject of gene editing accessible to other students, clarify open questions and create an active exchange of opinions. This idea is to be further pursued, as one member of the debating club wants to call more attention to the debate of gene editing inside the youth organisation of one of the large German parties (Social Democratic Party, SPD).

Scientific Commitment

In recent years, the relationship to the next generation of scientists has always been an important aspect of our public relations work in our team. Therefore, we continued our past successes this year and invited six high school students with their teachers and a father to our laboratory. We showed them standard methods of molecular biology, embedded in the background, to detect Bt corn in corn products they had brought with them. First, the DNA was isolated, purified and amplified with primers from Bt gene primers. The PCR products were then analysed in an agarose gel and the results discussed. Such experiments are an important part of the curriculum of secondary schools, as the equipment is usually not available on site. We deepen the current curriculum content, which is practically applied and thus no longer remains abstract. We overcome fears of contact with stigmatized genetic engineering. In addition, our invitation offers teachers a platform for further training in advanced teaching techniques that are up-to-date and applicable. The feedback on our test day was overwhelmingly positive, so we will continue to implement this format in future iGEM years.

Interview with Boris Palmer

Boehringer-Ingelheim

To see how we could possibly expand our project to the future, we organised a visit to Boehringer-Ingelheim which is one of the world’s top 20 pharmaceutical companies with expertise in diseases of the central nervous system, among others. We wanted to see how drugs are developed and produced and communicate with leading experts. Unfortunately, it was only possible to arrange a visit at the beginning of the next year.

Analytica Fair Munich

HyperpodX, University of Oldenburg

Luckily, we were able to cooperate with many student organisations inside Germany and various countries within Europe which enabled us to further reach out with our ideas, ambitions and the spirit of iGEM in general. In a meeting with the Hyperloop pod competition team of the University of Oldenburg (HyperpodX) both sides learned a lot about how to organize work and communication inside larger student groups and enjoyed the mutual exchange about two projects from very different areas.

German and European Meet Up

During the German Meet Up in Marburg and the European Meet Up in Munich we had great fun in presenting our project to other iGEM teams, exchanging experiences associated with the progress of our work and listening to intriguing talks.

Meeting iGEM Würzburg

This year our team also had a special connection with another iGEM team: iGEM Würzburg. Two members of this teams, Vic and Annabel went to school together. Nowadays, Vic is a member of our team while Annabel is the initiator of the first iGEM team of Würzburg. Back in time at school they used to compete against each other with their skills and knowledge about biology - and this year they worked together as young scientists to make their footprint in synthetic biology together with their teams. As Vic took part in iGEM last year Annabel came in touch with the synbio competition and was inspired by this great opportunity for challenges besides normal studying. So Annabel took the initiative and built the very first iGEM Team of Würzburg. They had mentoring during long skype calls - beginning with basic questions: What's needed for iGEM? How is a team structured? Due to the advice to go to fairs like the Analytica for fundraising it was possible for Annabel to establish a good and working team. On Meetups like the German and the European Meetup the teams got to know each other and talked about problems within the teams or how to set the focus during the projekt work. Since our team in Tuebingen already mastered many years of igem participation and already overcame various difficulties the team members provided Würzburg with many helpful advice. It’s great to see, that Würzburg will also go to Boston with their first project and some good results in the backpack. As science competitions are also a great possibility for stabilizing networks she stayed in touch with Annabel Maisl who is this years team leader of iGEM Würzburg, participating in iGEM for the first time.

Meeting iGEM Oslo

Our meeting with the iGEM Team Oslo was a great opportunity for us to present our project and our ideas on an international level for the first time while getting important feedback. Of course we also took the chance to get to know the city and culture of Norway. The first day in Oslo was mainly used to introduce ourselves and to get to know the team in Oslo. They offered us a rich program of cultural and scientific exchange. On the second day of our trip we as well as the team from Oslo gave first presentations of our projects. Afterwards we exchanged constructive criticism regarding the projects and its presentations. During the lunch break we also had the opportunity to talk to the PIs and instructors who also gave us valuable tips. This was followed by a workshop with the communications officer where many factors were discussed such as ways of implementing public relations in the field of natural sciences and how to transfer them specifically to our project. This lead to the development of plenty of good ideas that were partially put into practice through projects like the Ract! Festival or our public debate. It was a weekend full of inspiration that created great memories.

Collaboration with iGEM Paris

We are especially proud of our bioinformatics collaboration with iGEM Team Paris: we were able to use our prediction tool BERT (for more information on that please see our bioinformatics page) on their project about therapeutic proteins providing them with information about ways to deimmunize their proteins and gaining external validation for our tool. Furthermore, we exchanged knowledge about machine learning as well as molecular simulations. We together are very happy to say that BERTs results are excellent. The iGEM Team Paris additionally offered their help for another molecular dynamics simulation, which sadly will not be finished before wiki freeze.

In closing, this year we stand out as a team that is excellently connected with other iGEM Teams, companies and public figures and is as present on social media as never before. Documenting and sharing our development and progress on Facebook, Instagram and Twitter we were able to reach iGEM Teams all over the world informing about our project, iGEM and synthetic biology in general.