Team:Lethbridge Canada

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Contents

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.


This is our first year of participation and as such, we are pretty excited about the prospects. We are a group of students from the Chemical Engineering Department and from the School of Biosciences & Bioengineering. The most exciting aspect that we found about this competition was the interdisciplinary learning. A chemical reactor system invariably involves the design of control structures, and it is the design of these structures in a biological system that we wish to attain via our project.


A major objective 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 has used synthetic biology approaches to answer these questions. Our team comprises of nine undergraduates, three graduate students as student mentors and two faculty mentors, one each from biology and engineering background. The project specifically deals with the analysis of the effect of single and multiple feedback loops on gene expression. This project involves theoretical and experimental studies. We have designed synthetic constructs to mimic multiple feedbacks. The focus of our experimental work is to visualize the effect of multiple feedback loops on the synthetic construct using single cell analysis. The project provides insights into the roles of multiple feedback loops in biological systems.

Sponsors

Gold Sponsor: File:Iitbombaylogo.jpg Silver Sponsors: File:DSIR LOGO1.jpg

Team

Project

Diabetes
The purpose of our project is to create bacteria that will detect blood sugar levels and respond accordingly by producing insulin. Type one diabetes is caused by the degeneration of islet cells in the pancreas. Conventional methods of treatment for type one diabetes include direct injection of insulin intravenously, the transplantation of islet cells or even the introduction of an entirely new pancreas. Our engineered bacteria would provide a long-term solution compared to the standard injections, which need to be administered at least twice per day. In essence, our project has the potential to change the way that diabetes is treated.

Glucose Detection
For our project, a method of detecting glucose is necessary for our E. coli to initiate insulin transcription. If a change in blood sugar cannot be detected by the cell, then either insulin will not be produced or it would be overproduced. To engineer this, we plan to incorporate glucose transporter-2 (GLUT-2), already found in islet cells. Is a transmembrane protein that enables the rapid equilibrium of glucose concentrations on either side of the cell membrane. We will couple this with the use of a promoter that is sensitive to glucose (and possibly other sugars) and only active at a certain concentrations so the cells do not overproduce insulin at what would be considered a normal blood sugar level.

Insulin Production and Secretion
This glucose sensitive promoter will be coupled to the DNA for the production of the 51 amino acid polypeptide insulin (human insulin). Insulin needs to be produced and then exported out of the cell in order for the insulin to decrease the blood glucose levels. For the export of insulin, we plan to use a signal sequence that directs the cell to transport the insulin protein outside of the cell. This can be fused to the DNA that is in charge of the insulin production.

Delivery System
The method of delivery of our organism can be through direct delivery in the bloodstream or through the NASA Bio capsule. It is feasible to use a system that enters the blood stream because there are already millions of E. coli cells in our body, and we can use a strain that does not cause an immune response. Alternatively, the NASA Bio capsule, which is a tangle of carbon nanotubes that will be used to contain particular cells and eventually medicinal substances inside it, could also be used. The capsule could contain our cells, and, should the body need insulin, automatically start secretion. The Bio capsule is tiny, inserted into the skin, non-reactive and fast acting.

Notebook

March

Brainstorming

Project ideas:
Pests
· Bacterial Pest attractor (engineer bacteria to produce a substance, such as a pheromone or smell, that attracts and kills pests such as insects)
· Natural pesticides (engineer bacteria to produce a substance that repels or kills pests, such as those that harm crops, that can later be implemented into plants)
Water/Environment:
· Desalination of water (engineering bacteria to get rid of salt in salt water in order to make drinkable water from sea water)
· Getting rid of estrogen mimicking compounds in water (engineering bacteria to degrade them or sequester them)
· Waste treatment (engineering bacteria that can be integrated into waste water treatment)
· CFCs (engineer bacteria to produce metabolites that break down chlorofluorocarbons—compounds that contribute to the degradation of the ozone layer)
Health
· Stomach ulcers (creating a medication containing engineered bacteria to specifically target and kill Helicobacter pylori – the organism that causes stomach ulcers)
· Diabetes (engineered bacteria as Islet cells to produce insulin)
· Allergies/Immune system (engineer bacteria to produce antihistamines or alter epitopes)
· Bone density (engineer bacteria to produce and secrete calcium and other compounds to help heal broken bones or to prevent osteoporosis)
Process Improvement
· Oil fractionation catalyst (engineering bacteria to improve the separation of crude oil into valuable fraction and waste fraction)
· Nitrate fixation (engineer bacteria to improve nitrogen fixation so not as much fertilizer is needed)
Kill Switches (engineering bacteria to undergo induced or programmed cell death in order to control the organism)

April

May

June

Results/Conclusions

Safety

Attributions

Human Practices

Fun!