Difference between revisions of "Team:EPF Lausanne/Basic Part"
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<font size="100"><h3>Bikard et al. used dCas9-ω targetting the promoter PAM rich URS J23117, BBa_K1723001, in order to regulate gene expression using gRNA (single guide RNA) guided dCas9-ω. By using our own dCas9-ω system we proved that this promoter can be activated or repressed (see <a href="https://2015.igem.org/Team:EPF_Lausanne/Results">results page</a>). Now, on the model of this promoter, we created <b>a new fully synthetic promoter</b>: PAM rich URS J23117Alt promoter, BBa_K1723005. We mutated the sequence of BBa_K1723001 between and outside the -10 and -35 sequence where the RNA polymerase binds (see <a href="https://2015.igem.org/Team:EPF_Lausanne/Results">BBa_K1723005 registry page</a> for more details) in order to have <b>a promoter targeted by a set of sgRNAs different</b> from the sgRNAs aiming for BBa_K1723001. The creation of this part and its experimental validation (see <a href="https://2015.igem.org/Team:EPF_Lausanne/Results">results page</a>) is very promising for us as it is the proof of the MUTABILITY of the targeted promoters. We can now imagine of mutating again the promoter to obtain others promote/sgRNAs sets creating more and more different transistors-like elements in cells. </h3></font> | <font size="100"><h3>Bikard et al. used dCas9-ω targetting the promoter PAM rich URS J23117, BBa_K1723001, in order to regulate gene expression using gRNA (single guide RNA) guided dCas9-ω. By using our own dCas9-ω system we proved that this promoter can be activated or repressed (see <a href="https://2015.igem.org/Team:EPF_Lausanne/Results">results page</a>). Now, on the model of this promoter, we created <b>a new fully synthetic promoter</b>: PAM rich URS J23117Alt promoter, BBa_K1723005. We mutated the sequence of BBa_K1723001 between and outside the -10 and -35 sequence where the RNA polymerase binds (see <a href="https://2015.igem.org/Team:EPF_Lausanne/Results">BBa_K1723005 registry page</a> for more details) in order to have <b>a promoter targeted by a set of sgRNAs different</b> from the sgRNAs aiming for BBa_K1723001. The creation of this part and its experimental validation (see <a href="https://2015.igem.org/Team:EPF_Lausanne/Results">results page</a>) is very promising for us as it is the proof of the MUTABILITY of the targeted promoters. We can now imagine of mutating again the promoter to obtain others promote/sgRNAs sets creating more and more different transistors-like elements in cells. </h3></font> | ||
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Revision as of 01:49, 19 September 2015