"
Team:Paris Bettencourt/YonatanTest
From 2013.igem.org
| Line 81: | Line 81: | ||
</div> | </div> | ||
| - | + | ||
| + | |||
| + | <div id="page"> | ||
| + | <h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Target">Target</a></h2> | ||
| + | <div class="overbox"> | ||
| + | <div class="subbox1"> | ||
| + | <div class="bkgr"> | ||
| + | <h2>Background</h2> | ||
| + | <p>CRISPR/Cas systems generate site-specific double strand breaks and have recently been used for genome editing. </p> | ||
| + | </div> | ||
| + | <div class="aims"> | ||
| + | <h2>Aims</h2> | ||
| + | <p>Building a genotype sensor based on CRISPR/Cas that reports existance of an antibiotic resistance gene.</p> | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="subbox2"> | ||
| + | <div class="results"> | ||
| + | <h2>Results</h2> | ||
| + | <ul> | ||
| + | <li>Successfully cloned gRNA anti-KAN, crRNA anti-KAN, tracrRNA-Cas9 and pRecA-LacZ into Biobrick backbones and therefore generated four new BioBricks. </li> | ||
| + | <li>Testing the new assembly standard for our cloning.</li> | ||
| + | </ul> | ||
| + | </div> | ||
| + | <div class="biocriks"> | ||
| + | |||
| + | <center> <img width="95%" style="margin-top:30px" src="https://static.igem.org/mediawiki/2013/5/55/PB_Bb_target.png"/><br></center> | ||
| + | |||
| + | </div> | ||
| + | </div> | ||
| + | <div class="mainfig"> | ||
| + | <center> <img height="60%" src="https://static.igem.org/mediawiki/2013/3/31/PB_fig_target1.png"/></center> | ||
| + | <p style="margin-top:-60px;font-size:13px">CRISPR anti-Kan plasmids target kanamycin resistant E. coli. WT (blue) and a kanamycin resistant strain (KanR, red) were co-transformed with a plasmid carrying the Cas9 construct, and a second plasmid carrying the anti-Kanamycin gRNA. WT was successfully transformed with one or both plasmids. KanR E. coli couldn’t be tranformed with both plasmids because of Cas9-induced cleavage of the chromosome specifically at the KanR cassette, with about 99% efficiency. | ||
| + | </p> | ||
| + | </div> | ||
| + | </div> | ||
| + | <div style="clear: both;"></div> | ||
| + | |||
| + | </div> | ||
| + | |||
| + | |||
| + | |||
| + | |||
| + | |||
</html> | </html> | ||
<!-- ########## Don't edit below ########## --> | <!-- ########## Don't edit below ########## --> | ||
{{:Team:Paris_Bettencourt/footer}} | {{:Team:Paris_Bettencourt/footer}} | ||
Revision as of 12:34, 25 October 2013
Detect
Background
CRISPR/Cas systems generate site-specific double strand breaks and have recently been used for genome editing.
Aims
Building a genotype sensor based on CRISPR/Cas that reports existance of an antibiotic resistance gene.
Results
- Successfully cloned gRNA anti-KAN, crRNA anti-KAN, tracrRNA-Cas9 and pRecA-LacZ into Biobrick backbones and therefore generated four new BioBricks.
- Testing the new assembly standard for our cloning.
CRISPR anti-Kan plasmids target kanamycin resistant E. coli. WT (blue) and a kanamycin resistant strain (KanR, red) were co-transformed with a plasmid carrying the Cas9 construct, and a second plasmid carrying the anti-Kanamycin gRNA. WT was successfully transformed with one or both plasmids. KanR E. coli couldn’t be tranformed with both plasmids because of Cas9-induced cleavage of the chromosome specifically at the KanR cassette, with about 99% efficiency.
Target
Background
CRISPR/Cas systems generate site-specific double strand breaks and have recently been used for genome editing.
Aims
Building a genotype sensor based on CRISPR/Cas that reports existance of an antibiotic resistance gene.
Results
- Successfully cloned gRNA anti-KAN, crRNA anti-KAN, tracrRNA-Cas9 and pRecA-LacZ into Biobrick backbones and therefore generated four new BioBricks.
- Testing the new assembly standard for our cloning.
CRISPR anti-Kan plasmids target kanamycin resistant E. coli. WT (blue) and a kanamycin resistant strain (KanR, red) were co-transformed with a plasmid carrying the Cas9 construct, and a second plasmid carrying the anti-Kanamycin gRNA. WT was successfully transformed with one or both plasmids. KanR E. coli couldn’t be tranformed with both plasmids because of Cas9-induced cleavage of the chromosome specifically at the KanR cassette, with about 99% efficiency.
+33 1 44 41 25 22/25
team2013@igem-paris.org 
