Trondheim iGEM 2013

The project

All gram negative bacteria produce outer membrane vesicles (OMVs) by bulging of from their outer membrane. These OMVs contain different proteins and carry out varies functions such as quorum sensing, involvement in pathogenesis and transporting enzymes to distal locations. In our project we wish to prove that OMVs can be manipulated and to thereby implicate OMVs potential as a drug delivery vehicle. This is an innovative and novel approach to create a safe way to deliver drugs in the body.

We are going to prove that the OMVs can be manipulated by introducing two different proteins. The first one is a fusion protein of GFP and RFP. This will make it possible to visualize the vesicles as well as investigating the new functionalities of fusing two fluorescent proteins together. The second protein we wish to introduce to the vesicles is the transmembrane protein G derived from Streptococcus dysgalactiae ssp. equisimilisi. Protein G is known to bind to Human Serum Albumin (HSA) which helps S.dysgalactiae subsp. equisimilis hide from the immune system. Protein G will therefore be a potential important piece in a drug carrier by masking it from immunological destruction and making it stable in the blood stream.

In order to direct the proteins into the periplasm and vesicles they need to have a tat signal peptide at the N-terminal. This will transport the proteins through twin-arginine translocation pathway (Tat pathway). Studies show that a twin-arginine signal peptide is able to direct the export of active green fluorescent protein (GFP) in Escherichia coli . We also wish to be able to regulate the production and thereby the export of proteins to OMVs. In order to accomplish this task we will set the gene constructs under regulation of the Pm/Xyls promotor system. This is a positive regulation system that is activated by the inducer m-toluic acid

Introducing these proteins into vesicles also demonstrate that it is indeed possible to manipulate the content and therefore the properties of OMV's. We will prove the concept that OMVs can be engineered to meet the criteria of a drug delivery vehicle.

About us

This is the third year a team from the Norwegian University of Science and Technology (NTNU) is participating in the iGEM competition. Our team consists of five students with background in Biotechnology, Molecular Medicine and Medical Technology. Mostly our collective background is in molecular biology colored with biophysics, immunology and biotechnology. We have four advisors with different specialities! Rahmi is our lab-GOD and has all the answers. Eivind gives us money and inspiration, Martin is our GFP-expert and Gunvor with her past iGEM team experience has been a lot of help! We are naive students that want to come up with crazy ideas that supervisors are to experienced to consider an opportunity. Our project this year is HIGH-risk and it was almost immediately clear to us that this was what we wanted to do this year. Considering the time-limit of iGEM it was an ambitious project and had everything gone as planned and expected we could have reached our goal. We believe the project has potential and it should be further investigated. We've all enjoyed working as a team and learned tons about synthetic biology and the potential this field has feels unlimited! We have had a lot of fun, the life of a scientist is like a rollercoaster of ups and downs but the excitement and curiosity of research will always be a strong motivator!