Team:NAWI Graz/HP/Gold

Go to Top

At the beginning of our project, we participated a seminar called "Synthetic Biotechnology" supervised by our secondary PI. In the course of this lecture, we repeated the most used and popular cloning methods. The much more interesting part was learning some new cloning methods, which are on the cutting edge of gene technology. Subsequent to the seminar, we started to work out our project idea, finding a way to connect bacteria with a robot. At first, planning everything was really difficult, because we did not know how and where to begin. Nevertheless, we benefited from the long lasting experience of our secondary PI and got useful hints for designing of our constructs and how to start them to work in E. coli. Our secondary PI also suggested to work with the novel gene editing method CRISPR/Cas9. To get familiar with this system, we got offered an internship in the working group of CRISPR/Cas9.

Skype conference organized by University of Uppsala

Due to our project we took part in a skype conference about safety organized by the University of Uppsala. The discussion was about general safety in the lab, as well as the risks of the designed constructs and what might happen if they were released. To evaluate potential risks, we interviewed different experts about the safety aspects of our project and how to improve our constructs. However, we were able to enhance our designs according to their input and increased our knowledge in the field of genetic engineering.

Expert Interviews

Horizontal gene transfer, HGT, always represents a certain risk factor. In the course of the interviews, we evaluated, among other things, the safety assessment of our construct with various professors. The talks focused on the importance of incorporating deficiencies and discussing different options. Finally, we decided to use the following E. coli strains because of their deficiencies: BL21 DE3 Gold, XL1-blue, TOP10 and DH5alpha. After much discussion, we decided that these strains are the safest variant for our planned experiment.

Another problem we solved with the aid of an expert was the time delay from the production of the proteins till their degradation. After endless discussions, we came up with the solution to integrate a TEV-protease which accelerated the subsequent degradation.

Finally, for the fluorescence proteins a problem occurred referring the detection. Originally it was planned to detect the proteins after induction by means of a plate reader or through the robot, but since the fluorescence signal was way too low we had no chance in proving the presence of the proteins. Coincidentally, we came up with this problem during our discussion evening, which led to a lively discussion. Many solutions came up, finally we could agree that a Western Blot would be the most promising variant. And indeed, we have succeeded in detecting the protein. We are currently working on improving the fluorescence intensity. During Nerd Nite, we discussed possible improvements with dedicated students. Their intellectual approaches were very interesting, some of them have been implemented. The students were very enthusiastic that they were able to contribute to our project.

Some experts were able to support us with new input, in which areas our project could develop in the future. Many novel ideas that on how this project might emerge are listed in the application section. There, many speculative new ideas are described and some of our team members hope to be able to help bring them to life in the course of our scientific career.

Model

At the beginning of our project, we already had a model of our entire experimental process. Thus, we were able to take this to public events, such as the “Spritzerstandl”. There, we got a lot of input from the public and even made fundamentally changes on how the robot works. After fierce discussions, the pH-based controller has been improved by several aspects, instead of just reacting to sensors' left and right input signals, additionally a kind of memory has been introduced into the system through the creative use of the “bacterial medium”. By this, the robots could no longer get stuck in corners of the maze.

In addition, the model was sent to various other iGEM teams, who also created individually mazes. Unfortunately, we did not have time to solve this with our physical robot, but we can show in the model that this would theoretically be feasible.

Future iGEM Projects at Universities of Graz

Since donations to non-official associations can be difficult for many companies, we decided to found an official association called “NAWiGEM - association to support the projects of the Graz iGEM team” under the auspices of the DIY electro- and biohacker scene in Graz, the Realraum³ and the Open BioLab - Graz Austria.

To increase the degree of popularity of iGEM and everything connected to it, we promoted all iGEM related topics at our events. We constantly received new inquiries from interested students on how to participate in the competition. Through the interviews with various experts, we were able to increase our awareness in different institutes. This made it possible for us to appeal to students from different fields. The first meetings with the new participants already showed a huge interest of engineering students for the competition.