Difference between revisions of "Team:Bordeaux/Results"

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    <th class="tg-ytdm">Goals</th>
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    <td class="tg-4xo4">1. To study DSIP's effect on Caenorhabditis elegans
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    <br><br>2. To create a photo-inducible system controlling Caenorhabditis elegans
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    <br><br>3. To create a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic
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    <br><br>4. To add experimental results to the bio-informatics model</td>
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<h1><b>1. To study DSIP's effect on Caenorhabditis elegans</b></h1>
  
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<p align="justify">We built a plasmid containing DSIP under control of U6 promoter. This plasmid has been injected in several worms with another plasmid containing DsRed protein in order to compare injected worms to worms used as control. Worms have been observed by fluorescence microscopy.</p>
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<h4>&#x2794; No differences was observed between the two types of worms. DSIP doesn't seem to have any effect  on Caenorhabditis elegans.</h4>
  
 
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<h1>2. To create a photo-inducible system controlling Caenorhabditis elegans</h1>
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<p align="justify"><b>Inducer part</b> <br>
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We tried to create different constructions by changing the promoter used. </p>
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<ul> <li>Choosen promoters: pRPS-27, pXBP1, pUNC-119, pMYO2, pMYO3</li> </ul>
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<p align="justify">Except for pRPS27, all promoters own one or several  illegal restriction sites (White Collar-1 and White Collar-2  also own illegal restriction sites). However, pRPS27 amplification never worked and mutagenesis of others promoters was hard to do. So, we decided to work on an other promoter: U6.
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<br>
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Initial construction was to assemble WC1, P2A and WC2 under control of U6 promoter. By lack of time, a  simpler construction was realized: </p>
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<ul> <li>A first plasmid containing WC1 under control of U6 promoter,</li>
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<li>And a second plasmid containing WC2 under control of U6 promoter.</li></ul> </p>
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<p align="justify"><b>Induced part</b> <br>
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We created two constructions:
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<ul> <li>A first plasmid containing NLP-22 under control of FRQ promoter,</li>
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    <li>A second plasmid containing GFP under control of FRQ promoter.</li> </ul>
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This second construction permit to us to verify, thanks to GFP, that induced part works even if NLP-22 doesn't have any effect on Caenorhabditis elegans.
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</p>
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<h4>&#x2794; By lack of time, we didn't finish all of our constructions. So we were not able to validate the photo-inducible system. </h4>
  
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<h1>3. To create a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic</h1>
  
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<p align="justify">In progress</p>
  
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<p>Here you can describe the results of your project and your future plans. </p>
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<h1>4. To add experimental results to the bio-informatics model</h1>
  
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<h5>What should this page contain?</h5>
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<p align="justify">In progress</p>
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<li> Clearly and objectively describe the results of your work.</li>
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<li> Future plans for the project </li>
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<li> Considerations for replicating the experiments </li>
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    <th class="tg-1i7h">Results</th>
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    <th class="tg-ytdm">Perspectives<br></th>
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    <th class="tg-7lyn">Biobricks created in psB1C3<br></th>
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    <td class="tg-yw4l"><b>Sleep part</b><br><br>- No effect of DSIP on Caenorhabditis elegans<br>- Photo-inducible system not finished<br><br><b>Crispr part</b><br><br>- Crispr methylase not finished<br><br><b>Bio-informatics part</b><br><br>- In progress<br></td>
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    <td class="tg-4xo4"><b>Sleep part</b>                                                                                <br><br>- To finish photo-inducible system’s constructions<br>- To create better primers for pRPS-27 amplification<br><br><b>Crispr part</b><br><br>- To finish Crispr methylase constructions<br><br><b>Bio-informatics part</b><br><br>- To prepare a better protocol for bio-informatics experiments</td>
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    <td class="tg-baqh">pU6<br><br>pXBP1<br><br>pFRQ<br><br>NLP-22<br><br>pFRQ::GFP<br><br>P2A::GFP<br></td>
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<h5> Project Achievements </h5>
 
  
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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<li>A list of linked bullet points of the successful results during your project</li>
 
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
 
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Revision as of 17:57, 15 October 2016

Sleep with EpiC elegans

Goals
1. To study DSIP's effect on Caenorhabditis elegans

2. To create a photo-inducible system controlling Caenorhabditis elegans

3. To create a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic

4. To add experimental results to the bio-informatics model


1. To study DSIP's effect on Caenorhabditis elegans

We built a plasmid containing DSIP under control of U6 promoter. This plasmid has been injected in several worms with another plasmid containing DsRed protein in order to compare injected worms to worms used as control. Worms have been observed by fluorescence microscopy.

➔ No differences was observed between the two types of worms. DSIP doesn't seem to have any effect on Caenorhabditis elegans.

2. To create a photo-inducible system controlling Caenorhabditis elegans

Inducer part
We tried to create different constructions by changing the promoter used.

  • Choosen promoters: pRPS-27, pXBP1, pUNC-119, pMYO2, pMYO3

Except for pRPS27, all promoters own one or several illegal restriction sites (White Collar-1 and White Collar-2 also own illegal restriction sites). However, pRPS27 amplification never worked and mutagenesis of others promoters was hard to do. So, we decided to work on an other promoter: U6.
Initial construction was to assemble WC1, P2A and WC2 under control of U6 promoter. By lack of time, a simpler construction was realized:

  • A first plasmid containing WC1 under control of U6 promoter,
  • And a second plasmid containing WC2 under control of U6 promoter.

Induced part
We created two constructions:

  • A first plasmid containing NLP-22 under control of FRQ promoter,
  • A second plasmid containing GFP under control of FRQ promoter.
This second construction permit to us to verify, thanks to GFP, that induced part works even if NLP-22 doesn't have any effect on Caenorhabditis elegans.

➔ By lack of time, we didn't finish all of our constructions. So we were not able to validate the photo-inducible system.

3. To create a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic

In progress

4. To add experimental results to the bio-informatics model

In progress

Results Perspectives
 Biobricks created in psB1C3
Sleep part

- No effect of DSIP on Caenorhabditis elegans
- Photo-inducible system not finished

Crispr part

- Crispr methylase not finished

Bio-informatics part

- In progress
 Sleep part

- To finish photo-inducible system’s constructions
- To create better primers for pRPS-27 amplification

Crispr part

- To finish Crispr methylase constructions

Bio-informatics part

- To prepare a better protocol for bio-informatics experiments		 pU6

pXBP1

pFRQ

NLP-22

pFRQ::GFP

P2A::GFP

How to find us ?

Feel free to email us to provide some feedback on our project, have some information on the team and our work, or to just say hello !

Mail: igembdx@gmail.com
Copyright © iGEM Bordeaux 2016