Difference between revisions of "Team:TokyoTech/Experiment/Transcriptome Analysis"

 
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     <label for="vmcb-d"><a>Experiment</a></label>
 
     <label for="vmcb-d"><a>Experiment</a></label>
 
     <ul>
 
     <ul>
         <li style="padding-bottom: 10px; padding-top: 10px">
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         <li>
 
         <input type="checkbox" id="vmcb-d1" />
 
         <input type="checkbox" id="vmcb-d1" />
          <label for="vmcb-d1"><a style="text-align: center;">Bacteria <br>to Human Cells ▼</a></label>
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        <label for="vmcb-d1"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/Overview" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white" style="text-align: center;">Overview</a></label>
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        <input type="checkbox" id="vmcb-d2" />
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          <label for="vmcb-d2"><a style="text-align: center;">Bacteria to <br>Human Cells ▼</a></label>
 
             <ul>
 
             <ul>
 
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraI_Assay" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">TraI Assay</a></li>
 
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraI_Assay" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">TraI Assay</a></li>
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraI_Improvement" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">TraI Impovement <br>Assay</a></li>
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               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraI_Improvement" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">TraI Improvement <br>Assay</a></li>
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraR_Reporter_Assay" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white" >TraR Reporter <br> Assay</a></li>
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               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraR_Reporter_Assay" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white" >TraR Reporter <br>Assay</a></li>
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/Transcriptome_Analysis" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Transcriptome <br> Analysis</a></li>
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               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/Transcriptome_Analysis" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Transcriptome <br>Analysis</a></li>
 
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/Chimeric_Transcription_Factor" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Chimeric <br> Transcription <br> Factor Assay</a></li>
 
               <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Experiment/Chimeric_Transcription_Factor" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Chimeric <br> Transcription <br> Factor Assay</a></li>
 
             </ul>
 
             </ul>
 
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     <label for="vmcb-d2"><a style="text-align: center;">Human Cells to Bacteria ▼</a></label>
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     <label for="vmcb-d3"><a style="text-align: center;">Human Cells to <br>Bacteria ▼</a></label>
 
         <ul>
 
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           <li><a href="https://2017.igem.org/Team:TokyoTech/Experiment/AHK4_Assay" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">AHK4 Assay</a></li>
 
           <li><a href="https://2017.igem.org/Team:TokyoTech/Experiment/AHK4_Assay" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">AHK4 Assay</a></li>
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     <label for="vmcb-d3"><a href="https://2017.igem.org/Team:TokyoTech/InterLab" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white" style="text-align: center;">InterLab</a></label>
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     <label for="vmcb-d4"><a href="https://2017.igem.org/Team:TokyoTech/InterLab" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white" style="text-align: center;">InterLab</a></label>
 
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     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/HP" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Overview</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/HP" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Overview</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/HP/Silver" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Silver</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/HP/Silver" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Silver</a></li>
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/HP/Gold_Integrated" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Integrated <br> Human Practice</a></li>
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     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/HP/Gold_Integrated" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Gold (Integrated)</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Demonstrate" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Demonstrate</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Demonstrate" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Demonstrate</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Collaborations" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Collaborations</a></li>
 
     <li style="text-align: center;"><a href="https://2017.igem.org/Team:TokyoTech/Collaborations" onclick="w3_close()" class="w3-bar-item w3-button w3-hover-white">Collaborations</a></li>
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     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Introduction</b></h1><!-- 小見出し -->
 
     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Introduction</b></h1><!-- 小見出し -->
 
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    <p style="font-family: Poppins;font-size: 16px"><p style="text-indent:1em"> In our project, we worked on establishing an artificial inter-kingdom communication between human cells and bacteria. However, the inter-kingdom communication was not known well. Therefore,  we first had to choose appropriate signaling molecules to establish it. We chose one of the AHL molecules, 3OC8HSL, as a signaling molecule from bacteria to human cells. This is because it can interact with only genetically engineered human cells [1, read <a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraI_Assay">TraI Assay</a> page]. Next, we had to choose a signaling molecule from human cells to bacteria. Initially we had planned to use another AHL. However, we decided not to use AHLs as signal molecules from human cells to bacteria since AHLs cannot be synthesized by human cells due to lack of their materials [2]. Thus, we had to explore if there is a potential signaling molecule produced by human cells. To this end, <span style="font-style: italic">E. coli</span> cells were interacted with the supernatant of the medium which human cells were cultured. Then transcriptome analysis was conducted to observe if there were any differences in the gene expression in <span style="font-style: italic">E. coli</span>.</p>
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    <p style="font-size: 16px; text-indent: 1em"> In our project, we worked on establishing an artificial cross-kingdom communication between human cells and bacteria. However, the cross-kingdom communication was not known well. Therefore,  we first had to choose appropriate signaling molecules to establish it. We chose one of the AHL molecules, 3OC8HSL, as a signaling molecule from bacteria to human cells. This is because it can interact with only genetically engineered human cells (Read <a href="https://2017.igem.org/Team:TokyoTech/Experiment/TraI_Assay">TraI Assay</a> page). Next, we had to choose a signaling molecule from human cells to bacteria. Initially we had planned to use another AHL. However, we decided not to use AHLs as signal molecules from human cells to bacteria since AHLs cannot be synthesized by human cells due to lack of their materials [2]. Thus, we had to explore if there is a potential signaling molecule produced by human cells. To this end, <span style="font-style: italic">E. coli</span> cells were interacted with the supernatant of the medium which human cells were cultured. Then transcriptome analysis was conducted to observe if there were any differences in the gene expression in <span style="font-style: italic">E. coli</span>.</p>
  
 
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     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Summary</b></h1><!-- 小見出し -->
 
     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Summary</b></h1><!-- 小見出し -->
 
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    <p style="font-family: Poppins;font-size: 16px"><p style="text-indent:1em">We checked the difference of transcription between <span style="font-style: italic">E. coli</span> in normal medium and <span style="font-style: italic">E. coli</span> in the medium which human cells were cultured. Surprisingly, there were no significant differences in almost all of the RNA expressions except for  a couple of genes. The difference was only a little even for those genes. </p>
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        <p style="font-size: 16px; text-indent: 1em">We checked the difference of transcription between <span style="font-style: italic">E. coli</span> in normal medium and <span style="font-style: italic">E. coli</span> in the medium which human cells were cultured. Surprisingly, there were no significant differences in almost all of the RNA expressions except for  a couple of genes. The difference was only a little even for those genes. </p>
 
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    <p style="font-family: Poppins;font-size: 16px"><p style="text-indent:1em">This excel file is the data of the transcriptome analysis. The value of E- FDR of the all genes was 1.<center><a href="https://2017.igem.org/File:T--TokyoTech--Transcriptome_Analysis.xlsx">The result data of transcription analysis</a>
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        <p style="font-size: 16px; text-indent: 1em">The 4318 genes were checked and <span style="font-style: italic">malG</span>coding Maltose transport system permease protein MalG and <span style="font-style: italic">malF</span>coding Maltose transport system permease protein MalF were transcribed about 1.7-fold. This excel file is the data of the transcriptome analysis. The value of E- FDR of the all genes was 1.<center><a href="https://2017.igem.org/File:T--TokyoTech--Transcriptome_Analysis.xlsx">Click here to download the result data</a>
 
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     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Discussion</b></h1>
 
     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Discussion</b></h1>
 
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     <p style="font-family: Poppins;font-size: 16px"><p style="text-indent:1em">It said that the difference is statistically significant if the value of E-FDR is 0.5 or less.  Therefore, the results show that most of the genes were not activated or repressed by substances derived from human cells. Two genes related to maltose metabolism were transcribed about 1.7-fold higher. This fact indicates that a substance like maltose is secreted from human cells. However, it is not proper for a signaling molecule because the activation rate is very low. Considering this result, no substances derived from human can be used as our ideal signaling molecule. Therefore, we have to explore another substance which works as a signaling molecule (Read <a href="https://2017.igem.org/Team:TokyoTech/Experiment/AHK4_Assay">AHK4 Assay</a> page and Project page).
+
     <p style="font-size: 16px; text-indent: 1em">It is said that the difference is statistically significant if the value of E-FDR is 0.5 or less.  Therefore, the results showed that most of the genes were not activated or repressed by substances derived from human cells. Two genes related to maltose metabolism were transcribed about 1.7-fold higher. This fact indicates that a substance like maltose was secreted from human cells. However, it was not proper for a signaling molecule because the activation rate was very low. Considering this result, no substances derived from human could be used as our ideal signaling molecule. Therefore, we had to explore another substance which works as a signaling molecule (Read <a href="https://2017.igem.org/Team:TokyoTech/Experiment/AHK4_Assay">AHK4 Assay</a> page and Project page).
  
  
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     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Reference</b></h1>
 
     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Reference</b></h1>
 
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    <p style="font-family: Poppins;font-size: 16px"><p style="text-indent:1em">[1] Foundational Platform for Mammalian Synthetic Biology, 2012 Noah Davidsohn et.al
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        <p style="font-size: 16px; text-indent: 1em">[1] Foundational Platform for Mammalian Synthetic Biology, 2012 Noah Davidsohn et.al
    <p style="font-family: Poppins;font-size: 16px"><p style="text-indent:1em">[2] Multiple reference genomes and transcriptomes for <span style="font-style: italic">Arabidopsis thaliana</span>, 2011 Xiangchao Gan et.al
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        <p style="font-size: 16px; text-indent: 1em">[2] Multiple reference genomes and transcriptomes for <span style="font-style: italic">Arabidopsis thaliana</span>, 2011 Xiangchao Gan et.al
 
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<div class="w3-light-grey w3-container w3-padding-32" style="margin-top:75px;padding-right:58px"><p class="w3-right">Hajime Fujita:  <a href="96haji.me" title="W3.CSS" target="_blank" class="w3-hover-opacity">All Rights Reserved</a></p></div>
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<div class="w3-light-grey w3-container w3-padding-32" style="margin-top:75px;padding-right:58px"><p class="w3-right"><a href="http://96haji.me/" title="W3.CSS" target="_blank" class="w3-hover-opacity">Hajime Fujita with W3.CSS: All Rights Reserved</a></p></div>
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Latest revision as of 22:51, 1 November 2017

<!DOCTYPE html> Coli Sapiens

iGEM Tokyo Tech

Transcriptome Analysis


Introduction


In our project, we worked on establishing an artificial cross-kingdom communication between human cells and bacteria. However, the cross-kingdom communication was not known well. Therefore, we first had to choose appropriate signaling molecules to establish it. We chose one of the AHL molecules, 3OC8HSL, as a signaling molecule from bacteria to human cells. This is because it can interact with only genetically engineered human cells (Read TraI Assay page). Next, we had to choose a signaling molecule from human cells to bacteria. Initially we had planned to use another AHL. However, we decided not to use AHLs as signal molecules from human cells to bacteria since AHLs cannot be synthesized by human cells due to lack of their materials [2]. Thus, we had to explore if there is a potential signaling molecule produced by human cells. To this end, E. coli cells were interacted with the supernatant of the medium which human cells were cultured. Then transcriptome analysis was conducted to observe if there were any differences in the gene expression in E. coli.


Summary


We checked the difference of transcription between E. coli in normal medium and E. coli in the medium which human cells were cultured. Surprisingly, there were no significant differences in almost all of the RNA expressions except for a couple of genes. The difference was only a little even for those genes.


Results


The 4318 genes were checked and malGcoding Maltose transport system permease protein MalG and malFcoding Maltose transport system permease protein MalF were transcribed about 1.7-fold. This excel file is the data of the transcriptome analysis. The value of E- FDR of the all genes was 1.

Click here to download the result data


Discussion


It is said that the difference is statistically significant if the value of E-FDR is 0.5 or less. Therefore, the results showed that most of the genes were not activated or repressed by substances derived from human cells. Two genes related to maltose metabolism were transcribed about 1.7-fold higher. This fact indicates that a substance like maltose was secreted from human cells. However, it was not proper for a signaling molecule because the activation rate was very low. Considering this result, no substances derived from human could be used as our ideal signaling molecule. Therefore, we had to explore another substance which works as a signaling molecule (Read AHK4 Assay page and Project page).


Appendix: Material and Method



Reference


[1] Foundational Platform for Mammalian Synthetic Biology, 2012 Noah Davidsohn et.al

[2] Multiple reference genomes and transcriptomes for Arabidopsis thaliana, 2011 Xiangchao Gan et.al