Team:IIT Bombay India

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| Biological systems are known to have a great degree of regulation in their activity, and this higher level of control is attributed to the multiple levels of feedback that exist within any biochemical pathway. As a means of gaining deeper insight within the utility of multiple feedback loops, we have constructed 4 strains containing plasmids with modified lac operon. The lacI produced as a result of the expression can inhibit its own expression, causing one level of feedback, while it can also suppress the replication of plasmid, providing another level of control on the number of processes itself. By combination of these 2 controls, four different strains are possible. We wish to demonstrate the better control in the strain with multiple feedbacks as compared to the strain with no control by characterizing the inherent or stochastic error present in the system through simulations. Further, IPTG can bind to the lacI present in the system which would act as a repressor. Thus IPTG can act as an inducer for the system. The effects of varying concentrations of IPTG are also studied, the understanding being that a system with high IPTG concentration resembles that of an open loop system.
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| Established in 1958, IIT Bombay is one of the most recognized centers of academic excellence in the country today. The excellence of its academic programs, a robust  research and development program with  parallel improvement in facilities and infrastructure have kept it at par with the best institutions in the world. The ideas on which such institutes are built evolve and change with national aspirations, national perspectives, and global trends. At IIT Bombay we are continuously seeking to extend the boundaries of our research in a sustained manner with clear cut executable goals, grounded solidly in national realities.
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One of the major objectives of synthetic biology is to unveil the inherent design principles prevailing in biological circuits. Multiple feedback loops (having both positive and negative regulation) are highly prevalent in biological systems. The relevance of such a design in biological systems is unclear. Our team will use synthetic biology approaches to answer these questions. Our team comprises of nine undergraduates, 3 graduate students as student mentor and two faculty mentors, one each from biology and engineering background. The project specifically deals with the analysis of effect of single and multiple feedback loops on gene expression. This project will involve theoretical and experimental studies. We have designed synthetic constructs to mimic multiple feedbacks. The focus of our experimental work will be to visualize the effect of multiple feedback loops on the synthetic construct using single cell analysis. The project will provide insights into the roles of multiple feedback loops in biological systems.
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Further, any genetic regulation is manifested in the phenotype observed. Since lac operon is concerned with the successful utilization of lactose, we also characterize the growth of the system on lactose. The unrestrained expression of lacI represents a burden on the system, since lactose would be taken away by the existing lacI. Thus in the strain with multiple feedbacks, since it exhibits a greater control and reduction in noise for lacI expression, we also expect it to show greater growth with lesser error. Thus, ultimately, the genetic regulation achieved on the expression of lac operon is shown to ultimately control an observed phenotype, which is the culture growth.
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Revision as of 15:11, 21 October 2009

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Introduction

Established in 1958, IIT Bombay is one of the most recognized centers of academic excellence in the country today. The excellence of its academic programs, a robust research and development program with parallel improvement in facilities and infrastructure have kept it at par with the best institutions in the world. The ideas on which such institutes are built evolve and change with national aspirations, national perspectives, and global trends. At IIT Bombay we are continuously seeking to extend the boundaries of our research in a sustained manner with clear cut executable goals, grounded solidly in national realities.

One of the major objectives of synthetic biology is to unveil the inherent design principles prevailing in biological circuits. Multiple feedback loops (having both positive and negative regulation) are highly prevalent in biological systems. The relevance of such a design in biological systems is unclear. Our team will use synthetic biology approaches to answer these questions. Our team comprises of nine undergraduates, 3 graduate students as student mentor and two faculty mentors, one each from biology and engineering background. The project specifically deals with the analysis of effect of single and multiple feedback loops on gene expression. This project will involve theoretical and experimental studies. We have designed synthetic constructs to mimic multiple feedbacks. The focus of our experimental work will be to visualize the effect of multiple feedback loops on the synthetic construct using single cell analysis. The project will provide insights into the roles of multiple feedback loops in biological systems.

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