Difference between revisions of "Part:BBa K1941002:Experience"
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<strong><h5>Repression of TEF1 promoter</h5></strong> | <strong><h5>Repression of TEF1 promoter</h5></strong> | ||
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| + | [[File:T--EPFL--Repression TEF1 2PP7.png|200px|thumb|left|Repression_TEF1]] | ||
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| + | A) Repression design of Tef1 promoter. The hairpins 2PP7 recruit the two times RNA binding domain PCP fused with the transcriptional repressor protein Mxi1. Then, dCas9 carries the repressor complex to the promoter Tef1, guided by the recognition sequence of the scRNA. | ||
| + | B) Relative fluorescence histogram. GFP levels were measured using FACS after overnight cells growth. A bar graph shows fluorescence intensity of a strain transformed with three different plasmids carrying the following parts: Tef1-GFP, scTef1_2PP7, dCas9 and the fused repressor protein PCP-Mxi1 The data were normalized to report to the basal level: cells that have been integrated with a single plasmid carrying GFP under Tef1 promoter. A repression of 70 % GFP expression was noticeable. | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 22:42, 19 October 2016
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Applications of BBa_K1941002
Repression of TEF1 promoter
A) Repression design of Tef1 promoter. The hairpins 2PP7 recruit the two times RNA binding domain PCP fused with the transcriptional repressor protein Mxi1. Then, dCas9 carries the repressor complex to the promoter Tef1, guided by the recognition sequence of the scRNA. B) Relative fluorescence histogram. GFP levels were measured using FACS after overnight cells growth. A bar graph shows fluorescence intensity of a strain transformed with three different plasmids carrying the following parts: Tef1-GFP, scTef1_2PP7, dCas9 and the fused repressor protein PCP-Mxi1 The data were normalized to report to the basal level: cells that have been integrated with a single plasmid carrying GFP under Tef1 promoter. A repression of 70 % GFP expression was noticeable.
User Reviews
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