Team:CLS CLSG UK/Engineering
Credit - Bill Oxford@unsplash
Engineering
In order to achieve engineering success we have done a number of things: we have used and reused an iterative design model by which we were able to continuously improve our project. This led to us tackling and solving many of our projects problems using creative synthetic biology methods. As well as this, given we had very limited lab work, we were able to plan, in full, all of the lab work that we hoped to carry out, as well as plan and document the results obtained from our experiments.
Our iterative design process:
Throughout our Design it is clear that our process was a largely iterative one. We often had to go back to the drawing board late on in the project after new information came to light. A few exemplar examples of this are described below.
We first researched heavily into enzymes to use to best break down the cocaine and then settled on the bacterial cocaine esterase from rhodococcus. We then designed multiple genetic circuits that wld be able to produce the enzyme but settled on a simple but effective constitutive production. We then went on to build a model, with help from Imperial's team and then used our model to test the efficiency of our enzymes. From this we learnt that if we were able to increase the thermal half life of our enzyme, the ability to break down cocaine would be drastically increased. Having learnt this new information we then delved back into a mass of papers to try and use our newly learnt information. We discovered that there is a mutant form of our original enzyme with an increased thermal stability. We then went back and incorporated this enzyme into our original design. We have since uploaded both parts to the registry for future teams to build, test and learn with.
More generally though our whole project followed this rough design cycle:
Research
Our project began with much research into a variety of different fields and ideas. However, we soon realised that a project involving our local area would be most beneficial. Our eyes immediately turned towards the River Thames. While the River Thames is now considered one of the cleanest rivers in a major city, we knew there were still large issues. Some of our team members came across articles on the rising cocaine concentrations in the River Thames and the demise of the European Eel. This of course needed validation by scientific journals, but soon we realised that cocaine concentrations in the river Thames was having a large impact upon the European eel species. Please have a look at our description page for more about our research.
Design
We designed our project with all of our research in mind, we wanted to implement synthetic biology in a safe, efficient and sustainable manner to try and address this problem. Our design page details exactly how we achieved this. Our design ended up with three novel genetic circuits. As well as this we designed a new hardware device that is capable of getting in-situ, quantitative data that can be used by organisations, governments and other iGEM teams.
Build
We built various mathematical models that we could use to evaluate our system. As well as this, we built and adapted protocols so that, in the lab, we would be able to construct our circuits. Please see our methods and page for further detail.
Test and improve
We tested one of our constructs, the Synthetic Adhesin, the results of which can be seen here. As well as this, we used our model to test the use of different enzymes within our system to see their effectiveness. This can be seen here.
In addition, we planned out all our labwork, which included how we would approach and deal with problems. This can be seen here. While we were not able to complete all our labwork, we were able to test and show that one of our constructs works as expected, which is further detailed in our results.