Difference between revisions of "Team:Tongji Shanghai/Experiments"

 
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               <li><a href="#">Home</a></li>
 
               <li><a href="#">Home</a></li>
               <li>Project</li>
+
               <li>Team</li>
 
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+
             <div class="widget widget-categories" style="display: block;position: fixed;" id="sidebar1">
 
               <h4>Experiments<span class="head-line"></span></h4>
 
               <h4>Experiments<span class="head-line"></span></h4>
 
               <ul>
 
               <ul>
 
                 <li>
 
                 <li>
                   <a href="#">Topic 1</a>
+
                   <a href="#">Materials Synthesis</a>
 
                 </li>
 
                 </li>
 
                 <li>
 
                 <li>
                   <a href="#">Topic 2</a>
+
                   <a href="#">Plasmid Construction</a>
 
                 </li>
 
                 </li>
 
                 <li>
 
                 <li>
                   <a href="#">Topic 3</a>
+
                   <a href="#">Cell Experiments</a>
 +
                </li>
 +
                <li>
 +
                  <a href="#">Mice Experiments</a>
 
                 </li>
 
                 </li>
 
               </ul>
 
               </ul>
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            <div id="project">
+
            <!-- Classic Heading -->
+
           
+
              <h3 class="classic-title" style="margin-top:0px"><span><a  href="https://2015.igem.org/Team:Peking/Design/PC_Reporter">Paired dCas9 reporter</a></span></h3>
+
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+
                <div class="row">
+
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                    <img style="max-with:400px;" src="https://static.igem.org/mediawiki/2015/4/4e/Peking-Project-Overview1.png">
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+
                <p> It is well known that CRISPR/dCas9 has a unique ability to be programmed to bind any sequence with the assistance of sgRNA; it was conventionally used for DNA editing or genome study. In our project, however, we integrate split reporters into CRISPR/Cas9 by translationally fusing two fragments of a split reporter to dCas9, respectively, to convert the sequence-specific information of pathogenic bacteria's genome (in our case, <i>M. tuberculosis</i>) into easily readable signal including bioluminescence or pigment. We demonstrated that the PC reporter is highly compatible with NAD-based diagnosis using isolated genomic DNA of MTB.</p>
+
                </div>
+
              </div>
+
            </div>
+
  
              <h3 class="classic-title" style="margin-top:0px"><span><a  href="https://2015.igem.org/Team:Peking/Design/PC_Reporter">Paired dCas9 reporter</a></span></h3>
+
        <!-- Materials Synthesis -->  
              <div style="margin-top:0px; margin-bottom:0">
+
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                <div class="row">
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                  <div class="col-md-6">
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      <h3 class="classic-title" style="margin-top:0px"><span>Materials Synthesis</span></h3>
                    <img style="max-with:400px;" src="https://static.igem.org/mediawiki/2015/4/4e/Peking-Project-Overview1.png">
+
                  </div>
+
                  <div class="col-md-6">
+
                <p> It is well known that CRISPR/dCas9 has a unique ability to be programmed to bind any sequence with the assistance of sgRNA; it was conventionally used for DNA editing or genome study. In our project, however, we integrate split reporters into CRISPR/Cas9 by translationally fusing two fragments of a split reporter to dCas9, respectively, to convert the sequence-specific information of pathogenic bacteria's genome (in our case, <i>M. tuberculosis</i>) into easily readable signal including bioluminescence or pigment. We demonstrated that the PC reporter is highly compatible with NAD-based diagnosis using isolated genomic DNA of MTB.</p>
+
                </div>
+
              </div>
+
            </div>            
+
  
  
 +
 
 +
          <!-- Some Text -->
 +
            <ul style="margin-left:0px; font-size:16px">We have synthesized the AuNRs (gold nanorods) and detoxified them for photothermal therapy, also known as optical hyperthermia or photothermal ablation, which is an emerging strategy for treating solid tumors. Gold nanoparticles are capable of confining resonant photons, further inducing coherent surface plasmon oscillation of their conduction band electrons.<br><br>
 +
            For non-invasive therapy, near infrared (NIR) radiation is chosen because it penetrates tissue more deeply. And we find that AuNRs with a strong SPR(surface plasmon resonance) in the NIR region can show intense absorption of light in the NIR region, also biocompatibility. Even more, it can accumulate in tumor tissue via passive targeting phenomena. PEG can be used to increase biocompatibility, suppress immunogenic responses and decrease adsorption. </ul>
 +
          <br>
 +
          <br>
 +
         
 +
    </div>
 +
    </div>
 +
           
 +
        <!-- Cell Experiments-->
 +
        <div class="row">   
 +
        <div class="col-md-12" style="margin:0px; padding:0">
 +
            <h3 class="classic-title" style="margin-top:20px"><span>Plasmid Construction</span></h3> 
 +
          <!-- Some Text -->
 +
            <ul style="margin-left:0px; font-size:16px">Cancer thermotherapy realized by depositing heat into tumor in a minimally invasive way is a promising alternative to the conventional therapies for cancer treatment. It’s a therapeutic tool to eradicate cancer tumor with minimum toxic effects. To make it better, we plan to improve its ability to target to tumor cells and thermosensitivity of tumor cells. Furthermore, we hope it can reflect the the treatment process. Here, we provide an approach to optimize its tumor targeting and thermosensitivity of tumor cells by hTert promoter and heat shock protein 70(hsp70) prompter and the following tumor suppressor p53. Additionally, the luciferase following p53 makes it a reporter system.
 +
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 +
          <br>
 +
            <br>
 +
                   
 +
       
 +
         
 +
     
 +
         
 +
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 +
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 +
   
 +
   
 +
        <!-- Materials Synthesis -->
 +
    <div class="row">   
 +
      <div class="col-md-12" style="margin:0px; padding:0">
 +
      <h3 class="classic-title" style="margin-top:20px"><span>Cell Experiments</span></h3>
 +
      <!--<h4>Taipei Workshop in NYMU<span class="head-line"></span></h4><br>
  
 +
 
 +
          <!-- Some Text -->
 +
            <ul style="margin-left:0px; font-size:16px;">In order to test the toxicity and killing efficiency of our system, we have designed in vitro cytotoxicity test and in vitro photo thermal effect test. Cell viability is tested with cellTiter-Glo luminescent cell viability assay, which can reflect the ATP concentration of cells.<br><br>
 +
            </ul>
 +
          <br>
 +
          <br>
 +
         
 +
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 +
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+
   
 
+
    <div class="row">    
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+
      <div class="col-md-12" style="margin:0px; padding:0">
        </div>
+
      <h3 class="classic-title" style="margin-top:20px"><span>Mice Experiments</span></h3>
 
+
      <!--<h4>Taipei Workshop in NYMU<span class="head-line"></span></h4><br>
  
 +
 
 +
          <!-- Some Text -->
 +
            <ul style="margin-left:0px; font-size:16px;">Previous work has shown that golden nanorods is harmless to cells under 100μg/ml, and perform well when exposed to NIR. The feasibility of in vivo near-infrared OPTT is demonstrated after infected plasmids in tumor-bear mice by direct injection. Near-infrared OPTT was performed extra corporally using a portable wave diode laser.</ul>
 +
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 +
          <br>
 +
         
 +
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 +
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 +
   
 +
   
 +
   
 
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 +
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 +
     
 
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+
 
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Latest revision as of 19:12, 19 October 2016

Tongji_Shanghai-2016.igem.org Tongji Shanghai

Project

Our project is our story.

Materials Synthesis

    We have synthesized the AuNRs (gold nanorods) and detoxified them for photothermal therapy, also known as optical hyperthermia or photothermal ablation, which is an emerging strategy for treating solid tumors. Gold nanoparticles are capable of confining resonant photons, further inducing coherent surface plasmon oscillation of their conduction band electrons.

    For non-invasive therapy, near infrared (NIR) radiation is chosen because it penetrates tissue more deeply. And we find that AuNRs with a strong SPR(surface plasmon resonance) in the NIR region can show intense absorption of light in the NIR region, also biocompatibility. Even more, it can accumulate in tumor tissue via passive targeting phenomena. PEG can be used to increase biocompatibility, suppress immunogenic responses and decrease adsorption.


Plasmid Construction

    Cancer thermotherapy realized by depositing heat into tumor in a minimally invasive way is a promising alternative to the conventional therapies for cancer treatment. It’s a therapeutic tool to eradicate cancer tumor with minimum toxic effects. To make it better, we plan to improve its ability to target to tumor cells and thermosensitivity of tumor cells. Furthermore, we hope it can reflect the the treatment process. Here, we provide an approach to optimize its tumor targeting and thermosensitivity of tumor cells by hTert promoter and heat shock protein 70(hsp70) prompter and the following tumor suppressor p53. Additionally, the luciferase following p53 makes it a reporter system.


Cell Experiments

    In order to test the toxicity and killing efficiency of our system, we have designed in vitro cytotoxicity test and in vitro photo thermal effect test. Cell viability is tested with cellTiter-Glo luminescent cell viability assay, which can reflect the ATP concentration of cells.



Mice Experiments

    Previous work has shown that golden nanorods is harmless to cells under 100μg/ml, and perform well when exposed to NIR. The feasibility of in vivo near-infrared OPTT is demonstrated after infected plasmids in tumor-bear mice by direct injection. Near-infrared OPTT was performed extra corporally using a portable wave diode laser.