Difference between revisions of "Team:Bordeaux/Results"
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<td class="tg-4xo4"> | <td class="tg-4xo4"> | ||
<ol> | <ol> | ||
| − | <li>Studying DSIP's effect on Caenorhabditis elegans</li> | + | <li>Studying DSIP's effect on <i>Caenorhabditis elegans</i></li> |
| − | <li>Creating a photo-inducible system controlling Caenorhabditis elegans</li> | + | <li>Creating a photo-inducible system controlling <i>Caenorhabditis elegans</i></li> |
| − | <li>Creating a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic</li> | + | <li>Creating a Crispr/methylase in order to knock out sleep and allowing the study <i>Caenorhabditis elegans's</i> epigenetic</li> |
<li>Adding experimental results to the bioinformatics model</li> | <li>Adding experimental results to the bioinformatics model</li> | ||
</td> | </td> | ||
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</table> | </table> | ||
| − | <h3>1. Studying DSIP's effect on Caenorhabditis elegans</h3> | + | <h3>1. Studying DSIP's effect on <i>Caenorhabditis elegans</i></h3> |
<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> | <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> | ||
| − | <h5>➔ No differences was observed between the two types of worms. DSIP doesn't seem to have any effect on Caenorhabditis elegans.</h5> | + | <h5>➔ No differences was observed between the two types of worms. DSIP doesn't seem to have any effect on <i>Caenorhabditis elegans</i>.</h5> |
</div> | </div> | ||
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<div class="column half_size"> | <div class="column half_size"> | ||
| − | <h3>2. Creating a photo-inducible system controlling Caenorhabditis elegans</h3> | + | <h3>2. Creating a photo-inducible system controlling <i>Caenorhabditis elegans</i></h3> |
<h4 align="justify"><b>Inducer part</b></h4> | <h4 align="justify"><b>Inducer part</b></h4> | ||
<p align="justify">We tried to create different constructions by changing the promoter used. </p> | <p align="justify">We tried to create different constructions by changing the promoter used. </p> | ||
| − | <ul><li><u>Chosen promoters</u>: pRPS-27, pXBP1, pUNC-119, pMYO2, pMYO3</li> </ul> | + | <ul><li><u>Chosen promoters</u>: <b>pRPS-27, pXBP1, pUNC-119, pMYO2, pMYO3</b></li> </ul> |
<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: <b>U6</b>. <br> | <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: <b>U6</b>. <br> | ||
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<li>A first plasmid containing NLP-22 under control of FRQ promoter,</li> | <li>A first plasmid containing NLP-22 under control of FRQ promoter,</li> | ||
<li>A second plasmid containing GFP under control of FRQ promoter.</li> </ul> | <li>A second plasmid containing GFP under control of FRQ promoter.</li> </ul> | ||
| − | <p align="justify">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.</p> | + | <p align="justify">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 <i>Caenorhabditis elegans</i>.</p> |
<h5>➔ By lack of time, we didn't finish all of our constructions. So we were not able to validate the photo-inducible system. </h5> | <h5>➔ By lack of time, we didn't finish all of our constructions. So we were not able to validate the photo-inducible system. </h5> | ||
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<tr> | <tr> | ||
<td class="tg-yw4l"><b>Sleep part</b><br><br> | <td class="tg-yw4l"><b>Sleep part</b><br><br> | ||
| − | - No effect of DSIP on Caenorhabditis elegans<br> | + | - No effect of DSIP on <i>Caenorhabditis elegans</i><br> |
- Photo-inducible system not finished<br><br> | - Photo-inducible system not finished<br><br> | ||
<b>EpiCRISPR part</b><br><br> | <b>EpiCRISPR part</b><br><br> | ||
Revision as of 16:24, 18 October 2016
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. We tried to create different constructions by changing the promoter used. 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. We created two constructions: 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. In progress In progress
Goals
1. Studying DSIP's effect on Caenorhabditis elegans
➔ No differences was observed between the two types of worms. DSIP doesn't seem to have any effect on Caenorhabditis elegans.
2. Creating a photo-inducible system controlling Caenorhabditis elegans
Inducer part
Initial construction was to assemble WC1, P2A and WC2 under control of U6 promoter. By lack of time, a simpler construction was realized:
Induced part
➔ By lack of time, we didn't finish all of our constructions. So we were not able to validate the photo-inducible system.
3. Creating a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic
4. Adding experimental results to the bio-informatics model
Results
Perspectives
Biobricks created in psB1C3
Sleep part
- No effect of DSIP on Caenorhabditis elegans
- Photo-inducible system not finished
EpiCRISPR part
- Crispr methylase not finished
Bioinformatics part
- In progressSleep part
- Finish photo-inducible system’s constructions
- Create better primers for pRPS-27 amplification
EpiCRISPR part
Finish CRISPR methylase constructions
Bio-informatics part
- Prepare a better protocol for bio-informatics experimentspU6
pXBP1
pFRQ
NLP-22
pFRQ::GFP
P2A::GFP
