Team:Calgary/Project/Achievements

From 2013.igem.org

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<h1>Achievements</h1>
<h1>Achievements</h1>
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<h2>For our <b>Sensor</b> we...</h2>
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<h2>For our <span class="blue">Sensor</span> we...</h2>
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<ul><li>Amended two TALEs from the registry and resubmitted these parts. We optimized the TALEs such that they can be expressed in <i> E. coli </i> by removing the eukaryotic kozak sequence present in the original parts, codon optimizing it for <i> E. coli</i> and added a his-6 tag such that this protein can be purified using affinity chromatography.</li>
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<ul>
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<li> We submitted two novel reporters: beta-lactamase and ferritin. We showed that these enzymes </li>
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<li>Designed a <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor><span class="blue">modular platform system</span></a> which allows the interchangeability of a DNA binding protein, creating the potential for multiple different DNA sensors to be developed from the framework we have submitted</li>
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<li> We characterized beta-lactamase as a colorimetric reporter using phenol red. We also showed that beta
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<li>Designed a <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Detector><span class="blue">DNA binding TALE</span></a> which binds specifically to regions of the Shiga toxin gene <i>stx2</i> which are conserved amongst a wide variety of pathogenic <i>E. coli</i></li>
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<li>  
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<li>Submitted parts for an <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Reporter/PrussianBlueFerritin><span class="blue">engineered ferritin gene</span></a>, a biological nanoparticle capable of both scaffolding and stabilizing proteins, as well as carrying out catalytic reactions to act as a reporter</li>
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<h2>For our <b>Human Practices</b> we...</h2>
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<li><a href=https://2013.igem.org/Team:Calgary/Project/Detector><span class="blue">Designed, constructed, expressed, and successfully</span></a> purified proteins for both the DNA sensing elements of our prototype as well as the scaffolding and reporter components of the system</li>
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<h2>For our <b>Informed Design</b> we...</h2>
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<li>We showed that our <https://2013.igem.org/Team:Calgary/Project/PostRegionals><span class="blue">sensing elements</span></a> bind DNA with specificity on nitrocellulose membrane
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<li> We used Informed Design: We designed our project with the cattle industries culture and needs first and foremost. </li>  
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<li>We characterized the <span class="blue">kinetic properties</span> of our <span class="blue">sensing elements</span>
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<li> Our project began with extensive consultation with key cattle industry stakeholders who explained the industries culture and how our technology could best fit into this it. We spoke with a rancher (Bob Church), feedlot operator (Chinook Feeders), a veterinarian (Feedlot Management Services), a meat processor (Cargill), and vaccine producer (Bioniche). </li>  
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<li>Characterized the <a href="https://2013.igem.org/Team:Calgary/Project/OurSensor/Reporter/PrussianBlueFerritin"><span class="blue">kinetic properties and ability</span></a> of the ferritin protein to act as a reporter</li>
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<li> We continued to consult with these industry stakeholders throughout the development of our FerriTALE system to ensure our technology would continue to meet the needs of the industry. Their industrial experience helped us to at many points during the summer redesign and pivot our system to better fit within the industry.</li>  
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<li>Characterized the ability of our <a href="https://2013.igem.org/Team:Calgary/Project/OurSensor/Reporter/PrussianBlueFerritin"><span class="blue">purified ferritin</span></a> produced from our construct to act as a reporter in solution as well as on a nitrocellulose membrane</li>
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<h2>For our <b>Safety</b> we...</h2>
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<li>Showed that the E coil and K coil interact using an <a href="https://2013.igem.org/Team:Calgary/Project/OurSensor/Linker"><span class="blue">immunoprecipitation assay</span></a>.
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<li> Safety was a paramount concern with FerriTALES construction and use.  We developed them as an in vitro protein based system that significantly reduced the risk of an organism escaping. The FerriTALE detection system is produced with engineered bacteria, but only the proteins required to form the FerriTALE are used in the test strip system.</li>  
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<li>Characterized the <a href="https://2013.igem.org/Team:Calgary/Project/OurSensor/Reporter/BetaLactamase"><span class="blue">kinetic properties and ability</a> of the &beta; lactamase to act as a reporter</li>
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<li> Assessment of Accuracy: False positives and negatives could have a profound effect on the economics of FerriTALEs use.  We have included both a positive and negative controls in our system to reduce these possible impacts.</li>  
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<li>Created multiple versions of a <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Prototype><span class="blue">prototype for our final system</span></a>, including a lysis device</li>
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<h2>For our <b>Collaboration</b> we...</h2>
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<li>Performed <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Modelling><span class="blue">modelling</span></a> to inform both the <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Modelling/QuantitativeModelling><span class="blue">quantitative inputs</span></a> and construction of our system, as well as <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Modelling/SpatialModelling><span class="blue">visual modelling</span></a> in order to communicate how our system works spatially</li>
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<li> We collaborated with the Paris Bettencourt iGEM Team in developing a database of biosensors made in the iGEM competitions.</li>
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<li><a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Modelling/SpatialModelling><span class="blue">3D printed components</span></a> of our system in order to better understand how our components would interact at a molecular basis</li>
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<h2>For our <b>Parts</b> we...</h2>
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<li>Informed by modelling, we used &beta;-lactamase as a reporter and were able to demonstrate that <a href=https://2013.igem.org/Team:Calgary/Project/PostRegionals><span class="blue">the TALE capture system works</span></a>
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<li>We built a <a href=https://2013.igem.org/Team:Calgary/Project/OurSensor/Modelling/QuantitativeModelling><span class="blue">Scilab model</span></a> with the kinetic constants we found during our biobrick characterization that was capable of determining how quickly our final system would generate a response.</li>
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</ul>
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<h2>For our <span class="orange">Human Practices</span> we...</h2>
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<ul>
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<li>Created a dialogue with <a href=https://2013.igem.org/Team:Calgary/Project/HumanPractices/InformedDesign><span class="orange">multiple industry experts</span></a> and potential users in the field where our product could be applied</li>
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<li>Visited all steps of the <a href=https://2013.igem.org/Team:Calgary/Project/HumanPractices/InformedDesign><span class="orange">processing chain</span></a> in order to better understand the culture and practices of the industry</li>
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<li>Used discussions with our stakeholders to inform the design of our final system from <a href=https://2013.igem.org/Team:Calgary/Project/HumanPractices/InformedDesign><span class="orange">inception towards completion</span></a> so that it puts industry culture and needs first and foremost</li>
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<li>Designed our system with <a href=https://2013.igem.org/Team:Calgary/Project/HumanPractices/Safety><span class="orange">considerations towards safety</span></a>, within our laboratory and in the implementation of the final prototype</li>
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<li>Thought extensively about the <a href=https://2013.igem.org/Team:Calgary/Project/HumanPractices/Safety><span class="orange">implications of our system</span></a> in terms of the profound impact on community health and economics if false positives and negatives were to occur, and have included both  positive and negative controls in our system to mitigate these implications</li>
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 +
</ul>
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 +
<h2>For our <span class="yellow">Collaboration</span> we...</h2>
 +
 
 +
<ul>
 +
 
 +
<li>Worked with Paris-Bettencourt in <a href=https://2013.igem.org/Team:Calgary/Project/Collaboration/BiosensorDatabase><span class="yellow">defining biosensors</span></a> for the iGEM community</li>
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<li>Reviewed all previous iGEM projects in order to categorize those that were biosensors based on a <a href=https://2013.igem.org/Team:Calgary/Project/Collaboration><span class="yellow">consensus definition</span></a> made with Paris-Bettencourt</li>
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<li>Consolidated this information into an easy to use <a href=https://2013.igem.org/Team:Calgary/Project/Collaboration/BiosensorDatabase><span class="yellow">database (SensiGEM)</span></a> in order to allow for the community to search all previous sensors to gain useful insight and find useful parts when developing future projects</li>
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<li>Worked with <a href=https://2013hs.igem.org/Team:Consort_Alberta><span class="yellow">Consort Alberta</span></a> over the summer in order to develop a system for detecting xylene contamination in soils</li>
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</ul>
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<h2>For our <span class="yellow">Outreach</span> we...</h2>
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 +
<ul>
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<li>Continued our collaboration with our local science centre, Telus Spark, in order to educate our community about synthetic biology and <a href=https://2013.igem.org/Team:Calgary/Outreach/TelusSpark><span class="yellow">create activities</span></a> which would bring the science to a level which would allow for education through something fun</li>
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<li>Brought basic principles of synthetic biology to our city as a whole through our participation in running activites at our cities <a href=https://2013.igem.org/Team:Calgary/Outreach/Beakerhead><span class="yellow">Beakernight event</span></a></li>
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<li>Gave the world an insight into the daily life of an iGEMer as well as sharing synthetic biology through <a href=https://2013.igem.org/Team:Calgary/Outreach/GenomeAlberta><span class="yellow">blog posts</span></a> written for Genome Alberta</li>
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<li>Helped to start Consort, Alberta's first high school iGEM team along with the <a  href=https://2013.igem.org/Team:Calgary_Entrepreneurial><span class="yellow">Calgary Entrepreneurial team</span></a>, and helped them to bring home the human practices award</li>
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<li>Held a Science Café event at <span class="yellow">Sir Winston Churchill High School</span></a> talking about out project and how students in high school can get involved with projects like iGEM at the college and high school levels.  We were asked to return for the schools Science Expo Information Night.</li>
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</ul>
</section>
</section>
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Latest revision as of 06:46, 6 November 2013

Achievements

For our Sensor we...

  • Designed a modular platform system which allows the interchangeability of a DNA binding protein, creating the potential for multiple different DNA sensors to be developed from the framework we have submitted
  • Designed a DNA binding TALE which binds specifically to regions of the Shiga toxin gene stx2 which are conserved amongst a wide variety of pathogenic E. coli
  • Submitted parts for an engineered ferritin gene, a biological nanoparticle capable of both scaffolding and stabilizing proteins, as well as carrying out catalytic reactions to act as a reporter
  • Designed, constructed, expressed, and successfully purified proteins for both the DNA sensing elements of our prototype as well as the scaffolding and reporter components of the system
  • We showed that our sensing elements bind DNA with specificity on nitrocellulose membrane
  • We characterized the kinetic properties of our sensing elements
  • Characterized the kinetic properties and ability of the ferritin protein to act as a reporter
  • Characterized the ability of our purified ferritin produced from our construct to act as a reporter in solution as well as on a nitrocellulose membrane
  • Showed that the E coil and K coil interact using an immunoprecipitation assay.
  • Characterized the kinetic properties and ability of the β lactamase to act as a reporter
  • Created multiple versions of a prototype for our final system, including a lysis device
  • Performed modelling to inform both the quantitative inputs and construction of our system, as well as visual modelling in order to communicate how our system works spatially
  • 3D printed components of our system in order to better understand how our components would interact at a molecular basis
  • Informed by modelling, we used β-lactamase as a reporter and were able to demonstrate that the TALE capture system works
  • We built a Scilab model with the kinetic constants we found during our biobrick characterization that was capable of determining how quickly our final system would generate a response.

For our Human Practices we...

  • Created a dialogue with multiple industry experts and potential users in the field where our product could be applied
  • Visited all steps of the processing chain in order to better understand the culture and practices of the industry
  • Used discussions with our stakeholders to inform the design of our final system from inception towards completion so that it puts industry culture and needs first and foremost
  • Designed our system with considerations towards safety, within our laboratory and in the implementation of the final prototype
  • Thought extensively about the implications of our system in terms of the profound impact on community health and economics if false positives and negatives were to occur, and have included both positive and negative controls in our system to mitigate these implications

For our Collaboration we...

  • Worked with Paris-Bettencourt in defining biosensors for the iGEM community
  • Reviewed all previous iGEM projects in order to categorize those that were biosensors based on a consensus definition made with Paris-Bettencourt
  • Consolidated this information into an easy to use database (SensiGEM) in order to allow for the community to search all previous sensors to gain useful insight and find useful parts when developing future projects
  • Worked with Consort Alberta over the summer in order to develop a system for detecting xylene contamination in soils

For our Outreach we...

  • Continued our collaboration with our local science centre, Telus Spark, in order to educate our community about synthetic biology and create activities which would bring the science to a level which would allow for education through something fun
  • Brought basic principles of synthetic biology to our city as a whole through our participation in running activites at our cities Beakernight event
  • Gave the world an insight into the daily life of an iGEMer as well as sharing synthetic biology through blog posts written for Genome Alberta
  • Helped to start Consort, Alberta's first high school iGEM team along with the Calgary Entrepreneurial team, and helped them to bring home the human practices award
  • Held a Science Café event at Sir Winston Churchill High School talking about out project and how students in high school can get involved with projects like iGEM at the college and high school levels. We were asked to return for the schools Science Expo Information Night.