Team:Westminster/Overview

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<li><a href="https://2012.igem.org/Team:Westminster" class="navlink">Home</a></li>
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<li><a href="https://2012.igem.org/Team:Westminster/Team" class="navlink">Team</a></li>
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<li class="submenu-button"><a href="https://2012.igem.org/Team:Westminster/Protocols"class="navlink">Protocols</a></li>
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<h1>Background</h>
<h1>Background</h>
<h2>The Cancer Stem Cell Theory</h2>
<h2>The Cancer Stem Cell Theory</h2>
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<p>Since the development of the cancer stem cell theory, scientists and media have been buzzing with excitement over what they claim could be a breakthrough in the way we view cancer. The cancer stem cell theory says that only a small group of cells in cancers (called cancer stem cells) possess stem cell like characteristics (like the ability to differentiate) and have the ability to form new tumors.  This theory has since been experimentally proved; and has shown that these cancer stem cells produce elevated levels of aldehyde dehydrogenase; an enzyme involved in the normal oxidative metabolism of alcohol in the body. </p>
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<p>Many tumours contain a sub-population of self-renewing and expanding stem cells known as cancer stem cells whose symmetric division result in tumour growth. These cancer stem cells can be identified using molecular determinants or markers which have properties such as self-renewal, clonogenicity, multipotentiality, and longevity. Recent studies have increasingly implied the importance of Aldehyde dehydrogenase enzyme as a marker for cancer stem cells owing to their ability to detoxify potential cytotoxins, thus making them resistant to chemotherapy. </p>
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<p>These cancer stem cells have also been implicated in recurrence; current therapies don’t eliminate them and since they are the cells with the tumor-regenerating abilities, they are able to form new tumors and cause recurrence. </p>
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<h2>Our Project</h2>
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<h1>Our Project</h1>
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<p>Our iGEM project taps into the cancer stem cell theory. We have set out to synthesise a genetic construct that can identify, isolate and destroy cancer stem cells. Three different genetic constructs which serve different purposes are to be synthesised. A number of isoforms of ALDH exist, and three of the most common occurring form in cancer will be isolated. The isoforms selected are ALDH1A1, ALDH1A3 and ALDH3A1. These isoforms show increased activity in recurrent forms of cancer. We have also identified ALDH2, which will be used as the control isoform, as this form is involved in ethanol metabolism and it will be easily induced by the addition of ethanol.  </p>
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<p>Our project therefore aims to make biological constructs that would identify these cancer stem cells based on their aldehyde dehydrogenase producing capability and eliminate them as well. To do this, we identified the sequence for four isoforms of the mammalian aldehyde dehydrogenase and created biobricks out of them.  </p>
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<p>The second part of our project was to create constructs to selectively isolate cancer stem cells in a range of cancer cell lines. This will provide a powerful tool for researchers in the field of cancer Biology. <p>
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<p>The second part of our project was to create constructs to selectively isolate them. To do this, we chose to use the Plug and Play system of assembly created by DTU-Denmark for the 2011 iGEM competition and then make our biobricks RFC-21 compatible according to the guidelines for iGEM submission. To achieve this plug and play system, we contacted DTU and had them send us some of the parts they had made last year, as they were not available in the Parts Registry.   </p>
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<h2>Assembly System</h2>
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<h2>Mammalian Expression</h2>
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<p>  We have opted to use the Plug and Play system of assembly created by DTU-Denmark for the 2011 iGEM competition. In addition, we have designed our final constructs to conform to biobrick standard parts. We feel that this could offer an opportunity to incorporate Plug-n-Play to the Biobrick system.<p>
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<p>Because cancer is a human disease, we decided to test our constructs in mammalian cells. One challenge we had was finding parts for mammalian expression. For this reason, we contacted the Serrano labs and had them send us their entire eukaryotic promoter database.   </p>
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<p> Due to the scarcity of available mammalian in the registry, we had to source parts from outside the parts registry. We were fortunate to obtain parts from DTU-Denmark which they had made in 2011. These parts were not available in the Parts Registry. We also contacted Serrano labs and were fortunate to receive some mammalian biobrick parts from them. </p>
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Latest revision as of 03:10, 27 September 2012

Overview

Background

The Cancer Stem Cell Theory

Many tumours contain a sub-population of self-renewing and expanding stem cells known as cancer stem cells whose symmetric division result in tumour growth. These cancer stem cells can be identified using molecular determinants or markers which have properties such as self-renewal, clonogenicity, multipotentiality, and longevity. Recent studies have increasingly implied the importance of Aldehyde dehydrogenase enzyme as a marker for cancer stem cells owing to their ability to detoxify potential cytotoxins, thus making them resistant to chemotherapy.

Our Project

Our iGEM project taps into the cancer stem cell theory. We have set out to synthesise a genetic construct that can identify, isolate and destroy cancer stem cells. Three different genetic constructs which serve different purposes are to be synthesised. A number of isoforms of ALDH exist, and three of the most common occurring form in cancer will be isolated. The isoforms selected are ALDH1A1, ALDH1A3 and ALDH3A1. These isoforms show increased activity in recurrent forms of cancer. We have also identified ALDH2, which will be used as the control isoform, as this form is involved in ethanol metabolism and it will be easily induced by the addition of ethanol.

The second part of our project was to create constructs to selectively isolate cancer stem cells in a range of cancer cell lines. This will provide a powerful tool for researchers in the field of cancer Biology.

Assembly System

We have opted to use the Plug and Play system of assembly created by DTU-Denmark for the 2011 iGEM competition. In addition, we have designed our final constructs to conform to biobrick standard parts. We feel that this could offer an opportunity to incorporate Plug-n-Play to the Biobrick system.

Due to the scarcity of available mammalian in the registry, we had to source parts from outside the parts registry. We were fortunate to obtain parts from DTU-Denmark which they had made in 2011. These parts were not available in the Parts Registry. We also contacted Serrano labs and were fortunate to receive some mammalian biobrick parts from them.

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