Team:IIT Madras/Design

Creating synthetic biological products and playing around with life is no child’s play hence, as aptly put by Voltaire and famously resonated on multiple occasions like in the blockbuster movie Spiderman, “With great power, comes great responsibility” . We realize that synthetic biology and the ability to manipulate biological systems is an exceptional power and on this platform of open source distribution of knowledge as provided by iGEM, it is important to use this tremendous power wisely.

Keeping this in mind, our team has designed novel genetic constructs that when used are agreeable to the environment, are safe, controllable, provide flexibility and yet, can be easily shared with and used by other enthusiasts of synthetic biology via this revolution in synthetic biology that is iGEM.

Our team project involves tackling the very complex problem of Shiga toxin by production of two very simple biomolecules – a 9 AA peptide sequence and a small biomolecule, indole-3-acetaldehyde. Both the deliverables of the project are environment friendly with no unforeseen ability to have any kind of negative impact.

The promoter system chosen for both the constructs is the pLuxR system so that the anti-toxin and the small molecule biofilm inhibitor are produced only in cattle rumen at the appropriate AHL concentration only. Theoretically, this allows endogenous control over the synthesis of final products in the cattle and practically, it allows us to precisely control how we produce our two potent biomolecules in the lab.

And finally, probably the most important aspects of our design is the flexibility that it offers in terms of sharing it with other synthetic biology enthusiasts and the ease with which they can modify our genetic constructs to meet their design requirements.

We have created a “Modular Plug-and-Play system” which simply means that we have designed our system in such a way that before every modular element used (RBS, Protein Coding Region and terminator) we have a restriction site before it so that individual parts can be isolated and different models of the same construct can be created based on the purposes of your experimentation. The introduction of restriction sites in the stuffer region between the RBS and Protein coding region for example allows you to choose and isolate these parts separately and design new constructs accordingly. A pictorial representation of the Plug-and-Play system has been given below for both the Gb3 construct and the I3A constructs and the use of restriction enzymes to create modular elements can be seen.

Using different restriction enzymes, the modular elements can be plugged and played accordingly.