Difference between revisions of "Part:BBa K1941001:Experience"
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===Applications of BBa_K1941001=== | ===Applications of BBa_K1941001=== | ||
<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_1PP7.png|200px|thumb|left|Repression_TEF1]] | ||
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| + | A) Repression design of Tef1 promoter. The hairpins PP7 the 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_PP7 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 with both constructs. | ||
| + | C) Flow cytometry data (LSRII Snoopy) to quantify the level of GFP expression under the control of Tef1 promoter. In light blue, a population of cells that express dCas9, scTef1_PP7 and PCP-Mxi1. In grey, a population that express basal level of GFP under the control of Tef1 promoter. | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 22:32, 19 October 2016
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Applications of BBa_K1941001
Repression of TEF1 promoter:
A) Repression design of Tef1 promoter. The hairpins PP7 the 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_PP7 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 with both constructs. C) Flow cytometry data (LSRII Snoopy) to quantify the level of GFP expression under the control of Tef1 promoter. In light blue, a population of cells that express dCas9, scTef1_PP7 and PCP-Mxi1. In grey, a population that express basal level of GFP under the control of Tef1 promoter.
User Reviews
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