Regulatory

Part:BBa_K1723001:Experience

Designed by: Emilie Cuillery   Group: iGEM15_EPF_Lausanne   (2015-09-08)


Applications of BBa_K1723001





EPF_Lausanne_transistor.png

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





Effects of of dCas9-ω (BBa_K1723000) on BBa_K1723001 promoter

EPF_Lausanne_RESULTS_EXPERIENCE.png

promoter strength was 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


This chart show that BBa_K1723000 can act as a transcription regulator on the promoter BBa_K1723001 by either activating or inhibiting it. We also see that when provided both gRNA Z4, activating, and gRNA Z35, inhibiting, BBa_K1723001's inhibition is stronger, then allowing the creation of our transistor-like elemens. Visit our website for more informations: http://2015.igem.org/Team:EPF_Lausanne




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