Team:Northwestern

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<p id = "project-description-title"> Project Description </p>  
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<p id = "project-description-title"> <!--<font size=1>N</font> <font size=2>N</font> <font size=3>N</font> <font size=4>N</font> <font size=5>N</font>--> NUtralize! </p>
<div id = "project-description">  
<div id = "project-description">  
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    This year the Northwestern iGEM team has decided to focus on oral health care as the theme of our project. It has been well characterized that lactic acid secreted by natural bacteria in the human mouth decreases pH, leading to cavity formation. The enamel demineralization threshold is known to be around pH 5.5. Previous research has found that the genes asr and gadA are highly expressed near pH 5.5 and below. Our project aims to take a two-pronged approach at combating the natural lactic acid secretion. We plan to create a promoter that is always on at a basal level as well as one that is induced to express at a high level as a result of low pH. This project will lay the foundation for a significant amount of research as the dual-state promoter that we develop could then be incorporated with a gene related to lactic acid catabolism or neutralization.  
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  <p>  Poor oral health (tooth decay and infection) can lead to many painful symptoms and complicated treatments. Mouth bacteria are a primary culprit in tooth decay, as lactic acid secreted by natural bacteria in the human mouth decreases pH, leading to cavity formation. The enamel demineralization threshold is known to be around pH 5.5.  The overall vision of our project is to prevent mouth pH from falling below 5.5 by engineering bacteria to sense pH and respond by increasing the alkalinity. To do this, we aimed to create genetic pH sensing elements that would work in the mouth. <p/>
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<p>Our project laid the foundation for a dual-state promoter consisting of two promoters in series: a constitutive promoter active at a (tunable) basal level and an upstream, pH-inducible promoter. Previous research has found that the E. coli genes Asr and GadA are highly expressed when pH drops below 5.5. This research drove us to choose the promoter elements for GadA and Asr as our pH-inducible promoters. </p>
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<p>We have characterized a new pH-responsive promoter (Asr) as well as an iGEM-characterized promoter (GadA) and a constitutive promoter (Lpp). Ongoing work will evaluate how promoters in series affect each other and further characterize the dual state phenomenon. Here we used E. coli  for the ease at which it is genetically manipulated to provide proof of principle. Additionally, we tested these promoters with GFP for an easy visual output. Ultimately the aim is to engineer bacteria native to the oral microbiome to sense and respond to drops in pH. We envision that these bacteria will induce enzymes that catalyze reactions to counteract the decrease in pH, preventing the demineralization of the tooth. </p>
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<div class = "first-row">  
<div class = "first-row">  
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<div id = "problem">  
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<p id = "first-row-title"> Summary of Problem </p>  
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<p id = "first-row-title"> Summary of Problem </p>
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<p id = "first-row-description"> Following any meals, the pH within the mouth usually drop substantially which can lead to cavities. </p>
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                <center> <a href = "https://2013.igem.org/Team:Northwestern/problem"> <img src="https://static.igem.org/mediawiki/2013/thumb/2/2c/Summary-of-problem.gif/600px-Summary-of-problem.gif" width="120" height="120"/> </a></center>
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<p id = "first-row-description"> Following meals, the pH within the mouth drops substantially which leads to cavities. </p>
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</div>
</div>
<div id = "pH">  
<div id = "pH">  
<p id = "first-row-title"> pH </p>   
<p id = "first-row-title"> pH </p>   
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                <center> <a href = "https://2013.igem.org/Team:Northwestern/detectpH"><img src="https://static.igem.org/mediawiki/2013/thumb/1/1e/Ph-image.gif/120px-Ph-image.gif" width="120" height="120"/></center>
<p id = "first-row-description"> The first step in combating cavities is to create a bacteria that can detect pH. </p>
<p id = "first-row-description"> The first step in combating cavities is to create a bacteria that can detect pH. </p>
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<div id = "dual-state">
<p id = "first-row-title"> Dual State Promoter </p>  
<p id = "first-row-title"> Dual State Promoter </p>  
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<p id = "first-row-description"> In order to be able to produce quick response in addition to a basal level, the team has designed the dual state promoter. </p>
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                <center> <a href = "https://2013.igem.org/Team:Northwestern/dualstate"><img src="https://static.igem.org/mediawiki/2013/thumb/1/14/Dual-state-image.gif/180px-Dual-state-image.gif" width="120" height="120"/></center> </a>
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<p id = "first-row-description"> In order for a quick response from basal level, we designed the dual state promoter. </p>
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Latest revision as of 04:04, 28 September 2013



NUtralize!

Poor oral health (tooth decay and infection) can lead to many painful symptoms and complicated treatments. Mouth bacteria are a primary culprit in tooth decay, as lactic acid secreted by natural bacteria in the human mouth decreases pH, leading to cavity formation. The enamel demineralization threshold is known to be around pH 5.5. The overall vision of our project is to prevent mouth pH from falling below 5.5 by engineering bacteria to sense pH and respond by increasing the alkalinity. To do this, we aimed to create genetic pH sensing elements that would work in the mouth.

Our project laid the foundation for a dual-state promoter consisting of two promoters in series: a constitutive promoter active at a (tunable) basal level and an upstream, pH-inducible promoter. Previous research has found that the E. coli genes Asr and GadA are highly expressed when pH drops below 5.5. This research drove us to choose the promoter elements for GadA and Asr as our pH-inducible promoters.

We have characterized a new pH-responsive promoter (Asr) as well as an iGEM-characterized promoter (GadA) and a constitutive promoter (Lpp). Ongoing work will evaluate how promoters in series affect each other and further characterize the dual state phenomenon. Here we used E. coli for the ease at which it is genetically manipulated to provide proof of principle. Additionally, we tested these promoters with GFP for an easy visual output. Ultimately the aim is to engineer bacteria native to the oral microbiome to sense and respond to drops in pH. We envision that these bacteria will induce enzymes that catalyze reactions to counteract the decrease in pH, preventing the demineralization of the tooth.

Summary of Problem

Following meals, the pH within the mouth drops substantially which leads to cavities.

Dual State Promoter

In order for a quick response from basal level, we designed the dual state promoter.

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