Difference between revisions of "Team:Utrecht"

 
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<a class="active" href=""https://2018.igem.org/Team:Utrecht"">Home</a>
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<a href="https://2018.igem.org/Team:Utrecht/AboutUs">About Us</a>
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<h1> Project Description </h1>
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<p>Bacteria need to respond to a wide range of chemical substances which can be harmful or          beneficial to them. They use sensory proteins that bind and respond to different chemical substances. These proteins are modular and consist of domains that are highly conserved among different species. Several recombinant proteins have been made in the past showing that their potential to be used as engineered biosensors.  Our team will create a living biosensor that can utilize different receptors to detect a broad range of chemicals through an adapted chemotaxis pathway by using bioluminescence. This new diagnostic laboratory tool is re-usable, innovative, inexpensive, flexible, and sustainable compared to current detection systems which often make use of
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monoclonal antibodies. This new diagnostic laboratory tool is applicable to different areas of research, including concentration-dependent detection of toxins which affect the environment, pathogens, disease states, and pesticides or chemicals affecting agriculture.</p>
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<p>Chemotaxis in <i>E. coli</i> is mediated by a highly conserved, specific, and well-studied  pathway. It consists of a two-component system, in which a receptor activates the kinase autophosphorylation activity of CheA. CheA subsequently donates its phosphoryl group to the response regulator protein CheY, which translocates to the flagellar motor where it interacts with Flim. This interaction results in a change of the rotational direction of the flagellum. As a consequence, the bacteria reorientates itself in a random way.<br>
 
  Our team will utilize the chemotaxis pathway to create a living sensor that can detect a wide range of chemical substances in a way that is cheaper, easier to use, and faster than the currently used antibody based detection methods. We will do this by expressing a kinase dependent split luciferase (KDSL) assay that measures the phosphorylation state of CheY, in an <i>E. coli</i> UU1250 strain that lacks all wild type chemotaxis receptors but expresses proof-of-principle custom made fusion receptors instead. This assay uses a heterodimerizing luciferase which only dimerizes upon binding of a ligand, by abrogating steric hindrance.<br></p>
 
  
<p>The assay consists of the N-terminal (N-luc2-416) and C-Terminal (C-luc398-550) halves of bacterial luciferase fused to a short linker. This linker consists of CheZ196-214, and CheY53-67 flanked by two 5 amino acid long linker sequences (GGSGG). CheY53-67 contains the D57 residue that is phosphorylated by CheA when the receptor is stimulated due to the binding of a ligand. CheZ196-214 contains the CheY binding site of CheZ. Increased activity of CheA will phosphorylate the D57 residue of CheY53-67. This will allow CheZ196-214 to bind P- CheY53-67. This causes stearic hindrance which prevents the N-luc and C-luc halves of luciferase to reconstitute which abrogates luciferase activity.<br>  
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Welcome to our wiki page. We are <b>Team Utrecht 2018</b>, a very <b>diverse</b> team from <b>Utrecht University</b> in the Netherlands. Want to know more about our team?
  We will create two different types of chemotaxis receptors by fusing the ligand binding domains of the cytokinin receptor PcrK, and the epinephrin receptor QseC to the intracellular domain of the <i>E. coli</i> Tar receptor. We opt to use three different fusion points for both receptors based on previously published successful recombinant chemotaxis receptors <sup>[1]</sup>. We will modify these sensors so they can measure different concentrations of ligand by modifying the methyl accepting residues of the Tar methylation helixes, and expressing different levels of recombinant receptor.<br></p>
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<a href = "https://2018.igem.org/Team:Utrecht/Team"> Click Here. </a>
  
<p><b>If our proof-of-principle system shows successful concentration-dependent detection of chosen ligands, it will further pave the way for cheap, effective, and modular bacterial-based detection systems.</b></p>
 
 
 
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1. Bi, S., Pollard, A. M., Yang, Y., Jin, F., & Sourjik, V. (2016). Engineering hybrid chemotaxis receptors in bacteria. ACS synthetic biology, 5(9), 989-1001.
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<b>Water</b> is arguably our most <b> precious resource</b>.
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Unfortunately, increased use of <b>pharmaceuticals</b> including <b>antidepressants</b> and chemotherapeutic medications are contaminating this valuable resource.
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After use of these medications, <b>remnants</b> are <b>disposed</b> through the sewage systems and eventually <b>contaminate surface waters</b>, resulting in an increased <b>pollutant concentration</b> in the <b>environment</b>.
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These <b>pharmaceutical waste products</b> are still <b>biologically active</b> in vertebrates.</br> For example, even low concentrations of antidepressants pose a <b>threat to ecological systems</b>.
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We talked to <b>water treatment companies</b>, <b>specialists</b> and other <b>stakeholders</b> about how to <b>improve</b> current detection <b>methods</b>. Key <b>requirements</b> for our biosensor included <b>safety</b>, <b>accuracy</b>, <b>high measurement rates</b> and <b>low costs</b>.
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See more on <a href = "https://2018.igem.org/Team:Utrecht/Human_Practices"> Integrated Human Practices.</a>
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Based on the <b>input</b> of experts and stakeholders, we developed the <b>DeTaXion biosensor</b>. <b>DeTaXion</b> improves <b>identification</b> of <b>antidepressants</b> in water, based on the <b>chemotaxis pathway</b> of <i>E. coli.</i>
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For a more detailed discussion you can visit our  <a href = "https://2018.igem.org/Team:Utrecht/Description"> Project Description.</a>
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To achieve the <b>detection of antidepressants</b>, we set up a <b>three step</b> approach <a href = "https://2018.igem.org/Team:Utrecht/Design">Project Design</a>. The <b>chemotaxis pathway</b> of <i>Escherichia coli</i> was <b>adjusted</b> in such a manner that <i>E. coli</i> can detect antidepressants. Upon <b>recognition</b> of the antidepressant agents, the <b>light signal</b> produced by the <i>E. coli</i> changes. The <b>concentration range</b> to be detected was customized.
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Next, we performed these steps in the <b>lab</b>, which resulted into interesting <a href = "https://2018.igem.org/Team:Utrecht/Results">Findings</a>.</br></br>
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Including a successfully developed <b>platform</b> to test the functionality of the <b>hybrid receptors</b>.
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Since we wanted to <b>enlighten</b> others about <b>biotechnology</b>, <b>water safety</b> and <b>valorisation</b>, we engaged the public at several <b>outreach events</b>, including our <a href = "https://2018.igem.org/Team:Utrecht/Conference"> biotechnology conference </a> and <a href = "https://2018.igem.org/Team:Utrecht/Public_Engagement#Events">“Weekend of Science”</a>, establishing an <b>open discussion</b>. More about this topic can be found in <a href = "https://2018.igem.org/Team:Utrecht/Public_Engagement">Public Engagement</a>.
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Furthermore, we <a href = "https://2018.igem.org/Team:Utrecht/Collaborations">collaborated</a> with other iGEM teams to get <b>valuable insights.</b></div>
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We also had to find the means to fund our project.
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Luckily, some parties were willing to invest
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their time and capital in us.
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We are very grateful to our <a href = "https://2018.igem.org/Team:Utrecht/Sponsors">Sponsors</a> and other <a href = "https://2018.igem.org/Team:Utrecht/Attributions">Contributors</a>.
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A special thank you for your help!
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After a lot of late nights, long hours in the laboratory, insightful conversations with stakeholders, sweat (maybe some tears) and a massive amount of laughter and joy.
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</br></br>
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We, <b>Team Utrecht 2018</b>, proudly present you our <b>applied product:</b>
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</br></br>
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<a href = "https://2018.igem.org/Team:Utrecht/Applied_Design"><i style = "font-size: 1.6vw"><u><b>DeTaXion</a>: A biosensor to rapidly identify harmful pharmaceuticals in surface waters.</i>
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Latest revision as of 19:56, 6 December 2018

Hello You!

Welcome to our wiki page. We are Team Utrecht 2018, a very diverse team from Utrecht University in the Netherlands. Want to know more about our team?

Click Here.
Water is arguably our most precious resource. Unfortunately, increased use of pharmaceuticals including antidepressants and chemotherapeutic medications are contaminating this valuable resource.
After use of these medications, remnants are disposed through the sewage systems and eventually contaminate surface waters, resulting in an increased pollutant concentration in the environment.
These pharmaceutical waste products are still biologically active in vertebrates.
For example, even low concentrations of antidepressants pose a threat to ecological systems.
We talked to water treatment companies, specialists and other stakeholders about how to improve current detection methods. Key requirements for our biosensor included safety, accuracy, high measurement rates and low costs. See more on Integrated Human Practices.
Based on the input of experts and stakeholders, we developed the DeTaXion biosensor. DeTaXion improves identification of antidepressants in water, based on the chemotaxis pathway of E. coli. For a more detailed discussion you can visit our Project Description.
To achieve the detection of antidepressants, we set up a three step approach Project Design. The chemotaxis pathway of Escherichia coli was adjusted in such a manner that E. coli can detect antidepressants. Upon recognition of the antidepressant agents, the light signal produced by the E. coli changes. The concentration range to be detected was customized.
Next, we performed these steps in the lab, which resulted into interesting Findings.

Including a successfully developed platform to test the functionality of the hybrid receptors.
Since we wanted to enlighten others about biotechnology, water safety and valorisation, we engaged the public at several outreach events, including our biotechnology conference and “Weekend of Science”, establishing an open discussion. More about this topic can be found in Public Engagement.

Furthermore, we collaborated with other iGEM teams to get valuable insights.
We also had to find the means to fund our project. Luckily, some parties were willing to invest their time and capital in us.


We are very grateful to our Sponsors and other Contributors. A special thank you for your help!
After a lot of late nights, long hours in the laboratory, insightful conversations with stakeholders, sweat (maybe some tears) and a massive amount of laughter and joy.

We, Team Utrecht 2018, proudly present you our applied product:

DeTaXion: A biosensor to rapidly identify harmful pharmaceuticals in surface waters.