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What is the context of this research?

The barrier to exploiting OMVs is the lack of a standardized system for producing and customizing OMVs. A series of genetic variants are known to improve the production of vesicles, however, the quantity and size of OMVs generated by these variants is not known. The scientific community needs an optimized, standardised system for producing OMVs. To do so, we will compare the size and production of OMVs of multiple mutants and make this data available to the scientific community.

We will then use this data to create a strain of E. coli that overproduces customizable OMVs. Ultimately, by packaging target proteins into these OMVs we plan to adapt them to various applications such as drug delivery or bioremediation.

What is the significance of this project?

Currently, synthetic biologists engineer living bacterial cells for release into various environments in order to solve problems, such as cleaning up an oil spill. In this case, the bacteria are engineered to metabolize the oil. Releasing bacteria, however, poses biosafety risks as the bacteria and synthetically modified genes persist in the environment long after the function is required.

In comparison, OMVs have a limited lifetime, do not contain genetic material, and do not replicate. Yet they can still carry and release the necessary proteins and enzymes to carry out various useful functions. Thus, if harnessed, OMVs could become a widely used tool for the application of synthetic biology in replacement of whole bacterial cells.

What are the goals of the project?

For our iGEM project, we plan to do as following. The first step is to develop a simple assay to score OMV formation. We will be measuring OMV size, stability and rate of production. We would like to use this assay to compare the effect of a range of mutations, both on their own and in combination. Based on this data we will generate a strain of E. coli that is optimized for protein packaging and OMV production, suitable for future applications in synthetic biology. As such, we will be able to modify our OMV system to solve environmental or medical issues. One possibility is the packaging of PETase (an enzyme that breaks down PET plastic) into OMVs, for release into contaminated waterways.