Team:Dundee

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     <h1 style="color: #FFFFFF;">Dundee University</h1>
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         <center><h2>Introduction</h2></center>
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        The Dundee iGem team began this project with a series of meetings during which ideas were explored, and areas of interest were discussed to find a project focus that appealed to the team. After much debating, the area of healthcare was chosen, and the project goals were refined to concentrate on a method to counteract corruption of the natural gut flora balance by clostridium difficile (<em>C. diff.</em>). Individuals undergoing long term antibiotic treatment can suffer from pseudomembranous colitis caused by <em>C. diff.</em>, and the team's aim is to modify Escherichia coli (<em>E. coli</em>) to attack <em>C. diff. </em> in a manner that restores the natural balance of the gut flora.
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        <h2><img src="https://static.igem.org/mediawiki/2012/f/fb/Abstract.png"></h2>
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<b>Six, Lyse and Obliterate: a synthetic silver bullet against healthcare acquired infection.</b><br>
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Hospital acquired infections are a global problem. One example is <I>Clostridium difficile</I>, a bacterial pathogen that infects patients undergoing prolonged antibiotic treatment and results in pseudomembranous colitis, a potentially fatal gut infection. This project aimed to design a synthetic bacterium that would respond to <I>C. difficile</I> infection and kill the pathogen <i>in situ</i>. <I>Escherichia coli</I> was engineered to secrete an endolysin from a bacteriophage that would specifically attack the <I>C. difficile</I> cell wall. The endolysin was fused to the extracellular components of an engineered Type VI Secretion System from <i>Salmonella</i>, which itself comprised 13 different proteins. In addition, a synthetic ‘inflammation biosensor’ was developed, based on a two-component system from <i>Salmonella</i>, with the aim of restricting endolysin secretion to the diseased colon only. Mathematical modelling was used to assist in the development of the laboratory work and to investigate potential therapeutic strategies beyond the scope of the experimental programme.
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        <center><h2><em>C.Diff</em> Lysis</h2></center>
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        Recent research <a href="https://2012.igem.org/Team:Dundee/References">[1]</a> investigating <em>C. diff.</em> cell disruption identified a bacteriophage &Phi;CD27 which encodes an endolysin that can effectively lyse <em>C. diff.</em> and which can be expressed within <em>E.coli</em>. It is the team's intention to create <em>E.coli</em> cells with capability to express &Phi;CD27 endolysin which will be delivered via a secretion system that permits transference of the endolysin through penetrative cell wall contact.
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The team plan to introduce the &Phi;CD27 endolysin to the <em>C. diff.</em> cells within the intestinal tract via the creation of a type VI secretion system <a href="https://2012.igem.org/Team:Dundee/References">[2]</a> which will be expressed within the <em>E. coli</em> cells. By adopting the type VI secretion system to facilitate endolysin delivery, the team hope to provide a foundation for systems that employ targetted <em>C. diff.</em> treatment.
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        Micrograph depicting Gram-positive <em>C. diff</em> bacteria using a .1µm filter.<br />
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        Public Domain : Obtained from CDC image library (http://phil.cdc.gov)
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Latest revision as of 21:03, 26 September 2012


Six, Lyse and Obliterate: a synthetic silver bullet against healthcare acquired infection.
Hospital acquired infections are a global problem. One example is Clostridium difficile, a bacterial pathogen that infects patients undergoing prolonged antibiotic treatment and results in pseudomembranous colitis, a potentially fatal gut infection. This project aimed to design a synthetic bacterium that would respond to C. difficile infection and kill the pathogen in situ. Escherichia coli was engineered to secrete an endolysin from a bacteriophage that would specifically attack the C. difficile cell wall. The endolysin was fused to the extracellular components of an engineered Type VI Secretion System from Salmonella, which itself comprised 13 different proteins. In addition, a synthetic ‘inflammation biosensor’ was developed, based on a two-component system from Salmonella, with the aim of restricting endolysin secretion to the diseased colon only. Mathematical modelling was used to assist in the development of the laboratory work and to investigate potential therapeutic strategies beyond the scope of the experimental programme.




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