Team:Lethbridge Canada

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='''Project Description'''=
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==''Diabetes''==
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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.
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==Glucose Detection==
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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. GLUT2, also known as solute carrier family 2 (facilitated glucose transporter) or member 2 (SLC2A2), is a transmembrane carrier protein that allows passive glucose movement across cell membranes, which 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.
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==Insulin Production and Secretion==
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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.
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==Delivery System==
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada Home]
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The delivery system of our organism to detect glucose and produce insulin 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.
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada/The_Team The Team]
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada/The_Project The Project]
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada/Results Results]
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada/Human_Practices Human Practices]
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada/Notebook Notebook]
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!align="center"|[https://2012hs.igem.org/Team:Lethbridge_Canada/Safety Safety]
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== '''Introduction''' ==
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| Insulin is a hormone produced by pancreatic beta cells that causes cell membranes to become more permeable to glucose. It helps regulate blood sugar levels in the body; more is secreted as more sugar enters the bloodstream after a meal is consumed. Insulin and glucagon (another pancreatic hormone that increases blood sugar levels) are partners in the negative feedback loop that control the body’s blood sugar concentration.
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Diabetes mellitus is a disease that is characterized by abnormally high blood glucose concentrations. Animal cells need glucose for energy, and these cells absorb glucose from the bloodstream with the help of insulin. Diabetes mellitus occurs when the immune system attacks and destroys the pancreatic beta cells that produce insulin so not enough is produced (type 1) or when the body’s cells become resistant to the action of insulin (type 2).
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Our goal is to use the natural glucose detecting mechanism found in Escherichia coli and combine it with the human insulin producing gene to create a “prosthetic” pancreatic beta cell. The transcription of insulin will only begin when a change in glucose levels in the cell’s external environment stimulates a sensory transduction chain of events.There are two parts to our project:
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1) Construct 1 focuses on the glucose sensing and gene regulating mechanism. We will implement the global respressor Mlc in order to control the rate at which the insulin gene is transcribed.
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2) Construct 2 focuses on insulin production and release.
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=='''Attribution'''==
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These people and institutions were vital in making the Lethbridge High School iGEM experience both possible and enjoyable:
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University of Lethbridge - use of facilities
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Dr. Hans-Joachim Wieden - advising, arranging funding, and administration
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Kim Orr- advising, organizing, as well as planning the trip to Indiana for the Jamboree
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University of Lethbridge iGEM Team - guidance and assistance with laboratory procedures
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Isaac Ward - help in teaching students proper usage of the spectrophotometer
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Nathan Dawson - advising
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Alvin Lee- advising
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Ben Vuong- advising
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Justin Luu - advising
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Sutherland Dube - advising, guidance, arranging lab times, and keeping everyone on the same page
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== '''Sponsors''' ==
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Latest revision as of 09:23, 17 June 2012

Home.JPG
Home The Team The Project Results Human Practices Notebook Safety



Introduction

Insulin is a hormone produced by pancreatic beta cells that causes cell membranes to become more permeable to glucose. It helps regulate blood sugar levels in the body; more is secreted as more sugar enters the bloodstream after a meal is consumed. Insulin and glucagon (another pancreatic hormone that increases blood sugar levels) are partners in the negative feedback loop that control the body’s blood sugar concentration.


Diabetes mellitus is a disease that is characterized by abnormally high blood glucose concentrations. Animal cells need glucose for energy, and these cells absorb glucose from the bloodstream with the help of insulin. Diabetes mellitus occurs when the immune system attacks and destroys the pancreatic beta cells that produce insulin so not enough is produced (type 1) or when the body’s cells become resistant to the action of insulin (type 2).


Our goal is to use the natural glucose detecting mechanism found in Escherichia coli and combine it with the human insulin producing gene to create a “prosthetic” pancreatic beta cell. The transcription of insulin will only begin when a change in glucose levels in the cell’s external environment stimulates a sensory transduction chain of events.There are two parts to our project:

1) Construct 1 focuses on the glucose sensing and gene regulating mechanism. We will implement the global respressor Mlc in order to control the rate at which the insulin gene is transcribed.

2) Construct 2 focuses on insulin production and release.

Attribution

These people and institutions were vital in making the Lethbridge High School iGEM experience both possible and enjoyable:

University of Lethbridge - use of facilities

Dr. Hans-Joachim Wieden - advising, arranging funding, and administration

Kim Orr- advising, organizing, as well as planning the trip to Indiana for the Jamboree

University of Lethbridge iGEM Team - guidance and assistance with laboratory procedures

Isaac Ward - help in teaching students proper usage of the spectrophotometer

Nathan Dawson - advising

Alvin Lee- advising

Ben Vuong- advising

Justin Luu - advising

Sutherland Dube - advising, guidance, arranging lab times, and keeping everyone on the same page






Sponsors

Sponsors 00112.jpg