Difference between revisions of "Team:Bordeaux/Results"

Revision as of 16:27, 18 October 2016

Sleep with EpiC elegans

Goals
Studying DSIP's effect on Caenorhabditis elegans Creating a photo-inducible system controlling Caenorhabditis elegans Creating a Crispr/methylase in order to knock out sleep and allowing the study Caenorhabditis elegans's epigenetic Adding experimental results to the bioinformatics model

1. Studying 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. Creating a photo-inducible system controlling Caenorhabditis elegans

Inducer part

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

Chosen 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. Creating a Crispr/methylase in order to knock out sleep and allowing the study of Caenorhabditis elegans's epigenetic

In progress

4. Adding experimental results to the bio-informatics model