Team:Uppsala/Project

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<h2>Assembly Plan</h2>
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<p>The Bactisille is the future of microbial combat. In your body, in drinking-water, in important ecosystems, in food and in sewage plants harmful bacteria can fester and cause problems both for the individual and for society as a whole. The conventional way of combating these bacteria is with chemicals such as antibiotics that kills not only bad microbes, but the good ones as well. Harming the natural bacterial flora in the body and in nature can cause as much trouble as the bad bacteria that we intend to kill in the first place.
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<tr><td><img src="https://static.igem.org/mediawiki/2014/8/82/Whole_system_Uppsala14.png" Style="width: 100%;"></img></td></tr>
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<tr><td><p>The Bactissiles two-mode system. Our Bactissile will at first be in the <b>Tracking mode</b>, expressing alot of the chemotaxis controling gene cheZ, which will make it randomly searching for <i>Y. enterocolitica</i>. It will also produce a silencing sRNA, spot42_USP45, which will silence the expression of the Bactissiles weapon, colicin Fy.  
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Enter the Bactissile.
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When <i>Y. enterocolitica</i> enters the Bactissiles vicinity, the Bactissile will switch to its <b>Attack mode</b>. The Bactissile will start to produce its weapon, colicin Fy and secretes it towards <i>Y. enterocolitica</i>. After a certain threshold of colicin Fy concentration, <i>Y. enterocolitica</i> will die.</p></td></tr>
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The Bactissile kills one bacteria, and one alone.
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<h2>Project overview</h2>
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<p>The Bactissile is the future of microbial combat. In your body, in drinking-water, in important ecosystems, in food and in sewage plants harmful bacteria can fester and cause problems both for the individual and for society as a whole. The conventional way of combating these bacteria involves chemicals such as antibiotics that kills not only harmful microbes, but the beneficial ones as well. Disturbing the natural bacterial flora in the body and in nature may in fact  cause as much trouble as the harmful bacteria that we intend to kill in the first place.
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</p>
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<h3>Enter the Bactissile</h3>
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<p>The Bactissile tracks and kills one type of bacteria and leaves other species intact. By sensing that specific harmful bacteria, targeting it and killing it, the Bactissile is a modern weapon to fight bacteria in many different environments, without affecting the surroundings. Sensing, targeting and killing are the backbone of the Bactissile, but to prove it functional we needed to choose one of the many possible pathogens to target. In light of the numbers of deaths due to gut bacteria in the world, and the menacing issue of antibiotic resistance we decided to prove our concept by battling the gut pathogen <i>Yersinia enterocolitica</i>.
<br><br>
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By sensing that one harmful bacteria, targeting it and killing it, the Bactissile is a modern weapon to fight bacteria in many environment and systems, without harming the surroundings. Sensing, targeting and killing is the backbone of the Bactissile, but to prove it functional, we needed to chose one of the many possible pathogens to target. In light of the numbers of deaths due to gut bacteria in the world, and the menacing issue of antibiotic resistance we decided to prove our concept by battling the gut pathogen Yersina entercolitica.
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The natural approach to battling a harmful gut bacteria is through a beneficial gut bacteria. This is why we intended to introduce the Bactissile system in a probiotic, such as <i>Lactobacillus</i>, which last years Team from Uppsala worked with.</p>
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<h3>The pathogen <i>Yersinia enterocolitica</i></h3>
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<p><i>Y. enterocolitica</i> is a pathogenic bacteria related to <i>Y. pestis</i>, the cause of the Bubonic plague.  <i>Y. enterocolica</i> thrives in uncooked food and grows well even in refrigerated temperatures.  This pathogen is especially dangerous for young children, causing fever, diarrhea and severe dehydration. All pathogens causing food poisoning and diarrhea are particularly dangerous in underdeveloped countries, where hygiene, access to water and medical care is highly inaccessible.</p>
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<tr><td><p>The Bactissiles is in its <b>Tracking mode</b>, expressing alot of cheZ and sRNA, spot42_USP45.</p></td><td><img class="scheduleSmall" src="https://static.igem.org/mediawiki/2014/6/69/Tracking_new_Uppsala14.png" </img></td></tr>
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<h3>The Bactissiles two-mode system</h3>
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<p>We have constructed a two-mode system. When our Bactissile is far away from our target, <i>Y. enterocolitica</i>, the first mode will be active, the <b>Tracking mode</b>. This entails that our Bactissile’s production of cheZ and a silencing sRNA spot42_USP45 is induced by having the activator YenR interact with the recognition region, the yenbox, and thereby inducing the strength of the wildtype promoter fused to it. The increased amount of cheZ will result in that the Bactissile will take big leaps, randomly searching for <i>Y. enterocolitica</i>. Meanwhile the silencing sRNA will silence the translation and expression of the Bactissile’s weapon, the killing substance colicin Fy, by binding to its corresponding mRNA.
<br><br>
<br><br>
-
The natural approach to battling a bad gut bacteria is through a good gut bacteria, which is why we intended to apply the Bactissile system in a probiotic, such as Lactobacillus, which Team Uppsala 2013 worked with last year.  
+
Once <i>Y. enterocolitica</i> gets close to our Bactissile, <i>Y.enterocolitica’s</i> unique signaling molecule will start to enter our system and interact with our activator, YenR. When the signaling molecule binds to YenR, YenR will lose its active shape and thereby its ability to interact with the yenbox, and can no longer induce the expression of the promoter. When this happens, the amount of cheZ and silecing sRNA’s will decrease, resulting in that the Bactissile will stop moving and basically start tumbling in circles. Also, the amount of the silencing sRNAs will not be balanced with the mRNAs coding for the killing substance colicin Fy, which will lead to initiation of the production of its weapon, colicin Fy. These two processes is what we define as the trigger of the <b>Attack mode</b>.
<br><br>
<br><br>
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<h3><i>Yersinia entercolitica</i></h3>
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</p>
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<i>Y. entercolitica</i> is a pathogenic bacteria related to <i>Y. pestis</i>, the cause of the Bubonic plague. <i>Y. enterocolitica</i> thrives in uncooked food and grows well even in refrigerated temperatures. This pathogen is especially dangerous for young children, causing fever, diarrhea and severe dehydration. All pathogens causing food poisoning and diarrhea are particularly dangerous in underdeveloped countries, where hygiene, access to water and medical care is highly inaccessible.</p>
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<table Style="border-radius: 25px;">
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<tr><td><img class="scheduleSmall" src="https://static.igem.org/mediawiki/2014/0/0a/Attacking_new_Uppsala14.png" </img></td><td><p>The Bactissile have entered the <b>Attack mode</b> and have started to produce alot of its weapon, colicin Fy.</p></td></tr>
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Colicin Fy molecules will start being exported out of our host, interfering with the structure of <i>Y. enterocoliticas</i> membrane. When the concentration of colicin Fy secreted reaches the threshold of 1.4e10 molecules/mL the dosage will become lethal to <i>Y. enterocolitica</i>.
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<a href="https://2014.igem.org/Team:Uppsala/Project_Sensing"><img class="next_pic" src="https://static.igem.org/mediawiki/2014/d/dc/Uppsala-igem2014-Next.jpg"></a>
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Latest revision as of 21:25, 17 October 2014

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The Bactissiles two-mode system. Our Bactissile will at first be in the Tracking mode, expressing alot of the chemotaxis controling gene cheZ, which will make it randomly searching for Y. enterocolitica. It will also produce a silencing sRNA, spot42_USP45, which will silence the expression of the Bactissiles weapon, colicin Fy.

When Y. enterocolitica enters the Bactissiles vicinity, the Bactissile will switch to its Attack mode. The Bactissile will start to produce its weapon, colicin Fy and secretes it towards Y. enterocolitica. After a certain threshold of colicin Fy concentration, Y. enterocolitica will die.

Project overview

The Bactissile is the future of microbial combat. In your body, in drinking-water, in important ecosystems, in food and in sewage plants harmful bacteria can fester and cause problems both for the individual and for society as a whole. The conventional way of combating these bacteria involves chemicals such as antibiotics that kills not only harmful microbes, but the beneficial ones as well. Disturbing the natural bacterial flora in the body and in nature may in fact cause as much trouble as the harmful bacteria that we intend to kill in the first place.

Enter the Bactissile

The Bactissile tracks and kills one type of bacteria and leaves other species intact. By sensing that specific harmful bacteria, targeting it and killing it, the Bactissile is a modern weapon to fight bacteria in many different environments, without affecting the surroundings. Sensing, targeting and killing are the backbone of the Bactissile, but to prove it functional we needed to choose one of the many possible pathogens to target. In light of the numbers of deaths due to gut bacteria in the world, and the menacing issue of antibiotic resistance we decided to prove our concept by battling the gut pathogen Yersinia enterocolitica.

The natural approach to battling a harmful gut bacteria is through a beneficial gut bacteria. This is why we intended to introduce the Bactissile system in a probiotic, such as Lactobacillus, which last years Team from Uppsala worked with.

The pathogen Yersinia enterocolitica

Y. enterocolitica is a pathogenic bacteria related to Y. pestis, the cause of the Bubonic plague. Y. enterocolica thrives in uncooked food and grows well even in refrigerated temperatures. This pathogen is especially dangerous for young children, causing fever, diarrhea and severe dehydration. All pathogens causing food poisoning and diarrhea are particularly dangerous in underdeveloped countries, where hygiene, access to water and medical care is highly inaccessible.

The Bactissiles is in its Tracking mode, expressing alot of cheZ and sRNA, spot42_USP45.

The Bactissiles two-mode system

We have constructed a two-mode system. When our Bactissile is far away from our target, Y. enterocolitica, the first mode will be active, the Tracking mode. This entails that our Bactissile’s production of cheZ and a silencing sRNA spot42_USP45 is induced by having the activator YenR interact with the recognition region, the yenbox, and thereby inducing the strength of the wildtype promoter fused to it. The increased amount of cheZ will result in that the Bactissile will take big leaps, randomly searching for Y. enterocolitica. Meanwhile the silencing sRNA will silence the translation and expression of the Bactissile’s weapon, the killing substance colicin Fy, by binding to its corresponding mRNA.

Once Y. enterocolitica gets close to our Bactissile, Y.enterocolitica’s unique signaling molecule will start to enter our system and interact with our activator, YenR. When the signaling molecule binds to YenR, YenR will lose its active shape and thereby its ability to interact with the yenbox, and can no longer induce the expression of the promoter. When this happens, the amount of cheZ and silecing sRNA’s will decrease, resulting in that the Bactissile will stop moving and basically start tumbling in circles. Also, the amount of the silencing sRNAs will not be balanced with the mRNAs coding for the killing substance colicin Fy, which will lead to initiation of the production of its weapon, colicin Fy. These two processes is what we define as the trigger of the Attack mode.

The Bactissile have entered the Attack mode and have started to produce alot of its weapon, colicin Fy.

Colicin Fy molecules will start being exported out of our host, interfering with the structure of Y. enterocoliticas membrane. When the concentration of colicin Fy secreted reaches the threshold of 1.4e10 molecules/mL the dosage will become lethal to Y. enterocolitica.