"

From 2011.igem.org

DISColi:Bio-photolithography in Device Engineering Using Different Wavelengths of Light

The DISColi project aims to design and construct a novel bio-photolithographic system for the engineering of biofilms into functional 2D and 3D structures and devices in response to different patterns and wavelengths of light

In this project we worked with light responsive promoters, a novel biofilm-forming synthetic biology chassis, E. coli Nissle 1917, and novel biobricks including several designed for biofilm dispersal and fluorescent reporters with wider utility than GFP.

The main aims of our project can be separated into three light-controlled components: the designed sculpting of biofilms; 3D printing for encapsulation of cells; and the controlled modular synthesis of a variety of products.

Applications

Build your own nano-biofactory with modular product synthesis pathways - Use the DISColi system to form a biofilm then use different colours of light to select from a range cellular products from a single strain. Make an array of functionally diverse products from a common precursor, the products you want will only be in the biofilm cells (not the whole culture) allows for easy purification of the products in the amount you require.

Personalised nutritional supplements and and pharmaceuticals IN SPACE! - In remote locations which are difficult/costly to resupply DISColi system can be used. This means that a single culture can selectively product a range of useful compounds from a common precursor, such as opioids and isopenoids, tailored to exactly what is needed.

Microfluidics - Grow a microfluidics platform using biofilm to form the channels. These channels can be dispersed resetting the system and allowing the formation of new channels. Or alter the channels during experiments to create dynamic environments.

Tissue Engineering - Use precises laser light to form tiny 3D structures out of biofilm that can be used as a scaffold for tissue culture.

Light Controlled Multicellularity - Use different colours of light to control dispersal of different species of microbes allowing the precise construction of a mixed community biofilm.

Clean Bioreactors - Biofilms are costly to clean from pipes and bioreactors. Using the DISColi system of light based dispersal of biofilms can stop the build up and even break down excisting biofilms in pipes and bioreactors. Simply shine light of the right colour on your culture before you drain the vessel to trigger the dispersal mechanism will start production surfactant proteins that have been shown to disperse biofilm.

Highlights!

In the course of our project we have created many noteworthy biobricks and have made a series of very interesting discoveries. Here are our personal highlights, including our favourite biobricks, our new chassis, and our public presence. Have a look!

Judging Criteria

In this section we explain why we deserve a gold medal in accordance with the iGEM judging criteria. If you have very little time, this may be exactly what you are looking for!

E.coli Nissle 1917

Our new transformable, non-pathogenic, biofilm-forming chassis!

LOV2 and iLOV Reporters

LOV2 and iLOV are our incredible new reporters. Not only are they smaller, florescene brighter and recover from photobleaching faster than GFP but it also functions in anaerobic conditions! Try tagging your favorite proteins.

c-di-GMP Phosphodiesterase

c-di-GMP Phosphodiesterase breaks down c-di-GMP, which is a secondary messenger which regulates many behaviours such as motility and biofilm formation. Over-expressing this phosphodiesterase should decrease the levels of c-di-GMP, increasing cellular motility and causing biofilm dispersal. c-di-GMP has many more functions making this biobrick useful in a wide range of applications.

Ranaspumin2

The surfactant protein ranspumin-2 comes from foam nests Túngara Frog (Engystomops pustulosus). In DISColi we use it to disperse biofilm however its application are much broader.

Latherin

Latherin is another surfactant protein although this one is isolated from horse sweat.

Multiple Cloning Site Biobrick

We were slowed down due to repeated rounds of restrictions and ligations to put multiple different coding regions with the same promoter, RBS and terminator combo. So we designed this handy multiple cloning site biobrick to speed up that process.

Sponsors

With many thanks to our generous sponsors, without whom this project would not have been possible.

References

1) Mackenzie et al., 2009. Ranaspumin-2: structure and function of a surfactant protein from the foam nests of a tropical frog. Biophysical Journal, 96, pp. 4984-4992.