Part:BBa_K1723005:Experience

Applications of BBa_K1723005

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 dCas9-ω BBa_K1723000 on BBa_K1723005 promoter

promoter strength was 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

This chart show that BBa_K1723000 can act as a transcription regulator on the promoter BBa_K1723005 by either activating or inhibiting it. It still have to be proved that in presence of both inhibiting and activating sgRNAs, inhibition is stronger and this promoter could be used to form a transistor-like element as wanted.

Comparison of basal levels of promoter BBa_K1723005 [B_alt] versus promoter BBa_K1723001 [B]

promoter strength was assessed using GFPmut2 reporter.

This chart show that mutating the sequence of BBa_K1723001 without touching the -10 sequence and -35 sequence to obtain BBa_K1723005 didn't seem to have affect the promoter strength. Visit our website for more informations about our results and the use of this system as a transistor-like element:http://2015.igem.org/Team:EPF_Lausanne

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