Difference between revisions of "Part:BBa K2100000:Experience"
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We characterized the synthetic ERE3 promoter in three cell lines: MCF-7, tHESC, and ISH. All cell lines have endogeneous Estrogen Receptor alpha. We analyzed data from cells induced with estradiol (E2) and uninduced as a control.
 
We characterized the synthetic ERE3 promoter in three cell lines: MCF-7, tHESC, and ISH. All cell lines have endogeneous Estrogen Receptor alpha. We analyzed data from cells induced with estradiol (E2) and uninduced as a control.
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Experiment in MCF-7:
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We transfected MCF-7 cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 50 nM E2.
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https://static.igem.org/mediawiki/2016/f/f5/T--MIT--pERE3MCF7.png
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The results show a  fold difference in yellow fluorescent output between the induced MCF-7 cells and the uninduced cells  with a vehicle control (0.4% EtOH) to account for the possible increase in proliferation cells undergo when being induced with E2 dissolved in ethanol.
  
  
 
Experiment in tHESC:
 
Experiment in tHESC:
  
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We transfected tHESC cells with 250ng of hEF1a-mKate as a transfection marker and pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 50 nM E2.  
+
We transfected tHESC cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 50 nM E2.  
  
 
http://2016.igem.org/File:T--MIT--bhandarkar_ere3thesc.png
 
http://2016.igem.org/File:T--MIT--bhandarkar_ere3thesc.png

Revision as of 22:59, 17 October 2016


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Applications of BBa_K2100000

We characterized the synthetic ERE3 promoter in three cell lines: MCF-7, tHESC, and ISH. All cell lines have endogeneous Estrogen Receptor alpha. We analyzed data from cells induced with estradiol (E2) and uninduced as a control.

Experiment in MCF-7:

We transfected MCF-7 cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 50 nM E2.

T--MIT--pERE3MCF7.png

The results show a fold difference in yellow fluorescent output between the induced MCF-7 cells and the uninduced cells with a vehicle control (0.4% EtOH) to account for the possible increase in proliferation cells undergo when being induced with E2 dissolved in ethanol.


Experiment in tHESC:

We transfected tHESC cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 50 nM E2.

http://2016.igem.org/File:T--MIT--bhandarkar_ere3thesc.png

The results show a 9 fold difference in yellow fluorescent output between the induced tHESC cells and the uninduced tHESC cells with a vehicle control (0.4% EtOH) to account for the possible increase in proliferation cells undergo when being induced with E2 dissolved in ethanol.

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