Team:IIT Madras/Project

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== '''Overall project''' ==
== '''Overall project''' ==
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Our project is based on the fundamental concept of plasmid instability in a novel way to conceal information or ‘lock’ a gene’s function in a cell until the correct combination of inputs is fed into the cell. We call this a ‘combinatorial lock’. It involves the positive regulation of the gene of interest only on receiving the correct inputs from the user. We use plasmids which can confer resistance to certain antibiotics in the medium and link them up in a certain way (i.e, design a genetic circuit) so that they repress the expression of the gene of our interest. As the selection pressure is lifted from the media, the plasmids which have the repressors for the gene of interest are lost, hence revealing the gene on using the correct series of antibiotic washes. In essence, the process of unlocking would simply be the correct sequence of antibiotic media in which the cells should be washed. We would be working with both a 2 plasmid system as well as 3 plasmid system and it is easy to see that this principle, in theory, could be extended to n plasmids. We intend to demonstrate this with simulations in matlab. One can think of a number of potential applications of this system of ‘combinatorial locks’, an example being the field of medicine. Human body releases a myriad of chemicals everyday in a certain order according to external stimuli or otherwise. In case of a disease, if this order goes awry, then this lock can be used to release the required chemicals to bring back the desired equilibrium. Also, when one wants to have the expression of a gene of commercial interest available only to licensed users and not a third party, this lock could be used.
== Project Details==
== Project Details==

Revision as of 07:37, 22 August 2009

Example logo.png

Synthetic biology is a new area of biological research that combines science and engineering in order to design and build ("synthesize") novel biological functions and systems. It involves plugging together existing genetic components in order to create new and unique systems.

This application, titled the "Combinatorial Lock" introduces a new paradigm in gene regulation. The study is based on the concept of plasmid loss. Any episome introduced into the cell shows a segregational asymmetry accompanied with differential growth rates in the absence and presence of episome leading to an overall loss of the episomal unit in the absence of any maintaining selective pressure. It is hypothesized that by appropriately controlling the external selective pressures, we can thereby also control the direction of plasmid loss, modifying the existing gene regulation system in a pre determined manner. It is also hypothesized that introducing negative selective pressures against certain other directions of plasmid loss, in the form of constitutively repressed endotoxins will help streamline the regulatory system even further.

If successful, this study allows for exquisitely delicate and precise multifactorial control of gene regulation in the future. It is expected that if successful, this model can be used to also hide genes of commercial interest to protect it from unauthorized use.

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Contents

Overall project

Our project is based on the fundamental concept of plasmid instability in a novel way to conceal information or ‘lock’ a gene’s function in a cell until the correct combination of inputs is fed into the cell. We call this a ‘combinatorial lock’. It involves the positive regulation of the gene of interest only on receiving the correct inputs from the user. We use plasmids which can confer resistance to certain antibiotics in the medium and link them up in a certain way (i.e, design a genetic circuit) so that they repress the expression of the gene of our interest. As the selection pressure is lifted from the media, the plasmids which have the repressors for the gene of interest are lost, hence revealing the gene on using the correct series of antibiotic washes. In essence, the process of unlocking would simply be the correct sequence of antibiotic media in which the cells should be washed. We would be working with both a 2 plasmid system as well as 3 plasmid system and it is easy to see that this principle, in theory, could be extended to n plasmids. We intend to demonstrate this with simulations in matlab. One can think of a number of potential applications of this system of ‘combinatorial locks’, an example being the field of medicine. Human body releases a myriad of chemicals everyday in a certain order according to external stimuli or otherwise. In case of a disease, if this order goes awry, then this lock can be used to release the required chemicals to bring back the desired equilibrium. Also, when one wants to have the expression of a gene of commercial interest available only to licensed users and not a third party, this lock could be used.

Project Details

Part 2

The Experiments

Part 3

Results