Every year, a student team from the TU/e joins the iGEM competition. In 2017 the TU/e iGem team won a golden medal and was nominated for Best New Application!
The 2017 GUPPI project
The project included different proteins and an underlying concept seen in nature. In this section they will provide some additional information about the parts and principles on which their project is based.
Gelation and Liquid-Liquid Phase Separation
As the name of our project (GUPPI) states, we are making a system that initiates gell formation based multivalent protein protein interactions. In the design process we took inspiration from a common cellular organiser; Membraneless organelles.
Classical views on cellular organisation show that this is carried out by membrane surrounded organelles like the Golgi and Endoplasmic Reticulum. In recent years a lot has become clear about a new type of organizer, called membraneless organelles. These organelles are phase separated from their surrounding environment and function to separate or concentrate biological macromolecules.
Overview of generated results
During iGEM we generated many different sorts of results. In the beginning of iGEM we mostly worked on getting the right DNA plasmids, by using different methods, like Gibson assembly and Traditional Cloning. In this period, the most important results included agarose gels and sequencing result (they are presented in "DNA results"). After this period, we had to test the expression of our designed proteins and instead of agarose gels, SDS-PAGE gels became more important. The last period is marked by the "real" experiments, where we could finally test if our designed system indeed worked as planned. To get an overview of all the steps we performed in the lab, go see our "Notebook" page.
The results of our Proof of Concept can be found here.
As iGEM is a broad competition based around synthetic biology, we also did many other things, for example thinking about safety and the future, which are part of "Human Practices".
Another important part was making a model of our system. It took a lot of time to start up, but in the end we got to simulate our system ("Model Results"). During our collaboration with Potsdam we discovered that they had no team member with experience with modeling, so we offered to also model their system, the results can be found below our own results, but also on their WIKI page: "Potsdam"). Additionally, their lack of experience an the time it took for us to start up with the model, inspired us to design a software tool: "NFsim manual for dummies".
Accomplishments in short
- Successfully designed, expressed and characterized multivalent constructs that will gelate after an inducer is added.
- Modelled the behavior of GUPPI and made a general tool for simulating other constructs with the same principle.
- Contacted stakeholders and integrated their input for a safe and useful design of GUPPI.
- Uploaded sequences of parts, characterized expression in the pBAD vector and delivered a codon optimized mCherry part.
See our "Medal Criteria" page for a broad list of all our accomplishments during iGEM.