Part:BBa_K2100000:Experience

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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 5 nM E2.

The results show an 8 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.

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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