Coding

Part:BBa_K1723000:Experience

Designed by: Gregoire Thouvenin   Group: iGEM15_EPF_Lausanne   (2015-08-10)


Applications of BBa_K1723000




EPF_Lausanne_transistor.png

This picture represent the system we built using dCas9-ω : a transistor-like element in cell. The complex dCas9-sgRNA will bind on the promoter and activate or inhibit it.




Effects of BBa_K1723000 on BBa_K1723001 promoter

EPF_Lausanne_RESULTS_EXPERIENCE.png

promoters strength were assessed using GFPmut2 reporter.


This chart present the effects of dCas9-ω (BBa_K1723000) on the PAM rich URS J23117 promoter (BBa_K1723001) when provided following sgRNAs:

[A] sgRNA Z4 (BBa_K1723003), activating
[A/R1] sgRNA Z4 (BBa_K1723003), activating, AND sgRNA Z35 (BBa_K1723004), inhibiting
[A/R2] sgRNA Z4 (BBa_K1723003), activating AND sgRNA Z0 (BBa_K1723002), inhibiting
[R2] sgRNA Z0 (BBa_K1723002), inhibiting
[R1] sgRNA Z35 (BBa_K1723004), inhibiting

In comparison with:

[B] apo-dCas9-ω (=no sgRNA)
[Af] apo-dCas9-ω (=no sgRNA), no reporter




Effects of dCas9-ω BBa_K1723000 on BBa_K1723005 promoter

EPF_Lausanne_RESULTS_EXPERIENCE2.png

promoters strength were assessed using GFPmut2 reporter.


This chart present the effects of dCas9-ω (BBa_K1723000) on the PAM rich URS J23117Alt promoter (BBa_K1723005) when provided following sgRNAs:

[A_alt] sgRNA X4 (BBa_K1723007), activating
[R2_alt] sgRNA X0 (BBa_K1723006), inhibiting
[R1_alt] sgRNA X35 (BBa_K1723008), inhibiting

In comparison with:

[B_alt] apo-dCas9-ω (=no sgRNA)
[Af_alt] apo-dCas9-ω (=no sgRNA), no reporter


Those charts show that BBa_K1723000 can act as a transcription regulator on both promoters BBa_K1723001 and BBa_K1723005 by either activating or inhibiting them. By creating new promoters, this system can be extended. We also see that when provided both gRNA Z4, activating, and gRNA Z35, inhibiting, on the promoter BBa_K1723001's inhibition is stronger, then allowing the creation of transistor-like elements. Visit our website for more informations: http://2015.igem.org/Team:EPF_Lausanne




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