Team:Tuebingen/Human Practises
Human Practises
Let the beauty of what you love be what you do.- Rumi
Introduction
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!
Integrated Human Practises
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.
Public Engagement
We started this years Human Practises work with the goal in mind to significantly increase our outreach to the public. By being as present on social media as never before in our iGEM history and documenting and sharing our development and progress on Facebook, Instagram and Twitter along the way we were able to reach iGEM Teams all over the world informing them about our project, iGEM and synthetic biology in general.
The local “Ract!” festival with visitor numbers close to 30,000 was a great first opportunity to implement our goal which we realised by having an informational stand and an additional parcour that was highly frequented by people of all ages.
Our public debate in collaboration with our universities debating club Streitkultur e.V. about the topic of gene editing on embryos also turned out to be a huge success due to a very lively and interactive audience that received new insights and perspectives on this highly controversial topic.
Because of previous Human Practises projects with schools and their pupils it was dear to our hearts to keep up this tradition, and so we organised a special laboratory day for this next generation of young scientists that was filled with experiments around the detection of Bt corn in corn products.
Last but not least it was very important for us this year to put an emphasis on collaborating and meeting up with other iGEM teams on a national and international level. Whether it was through helping and mentoring the iGEM team Würzburg with the establishment of their team, collaborating with the iGEM team Paris on a very successful bioinformatic prediction tool (“BERT”), meeting up with iGEM team Oslo to present our project and exchange thoughts, advice and ideas, or attending big meet ups like the European or German Meet Up to get in contact with other teams, we are proud to have achieved and exceeded this goal.
If you would like to read more about our this years Public Engagement please visit the page linked below - thank you!