Team:MIT/Experiments/Promoters/Experiment-Details-for-Cascades

Estrogen Sensitive Promoters in Repressor Cacades

We began testing how our estrogen responsive promoters behave in larger genetic circuits by first testing estrogen repressor cascades. We considered three repressors, BM3R1, TAL14, TAL21.

Our Genetic Circuit for Repressor Cascade Characterization

Figure . Our estrogen sensitive promoters respond to increases in E2 levels by producing more of the repressor. The repressors then bind to binding sites in a promoter upstream of fluorescent reporter eYFP. The constitutively active trans-activator Gal4-VP16 sets a large basal eYFP expression when there is no repressor, so that a measurable drop in signal can be observed when repressors are active. Constituvely active hEF1a mKate serves as a transfection marker by which we bin our data.

Repressor Cascades in ISH

Figure . Results from pERE#n - repressor cascades pairing pERE3, pERE5, pERE6 and TAL14, BM3R1. Blue contours represent the cell population that was left uninduced, green contours represent the cell population that was induced with 5 nM E2.

Estrogen Sensitive Promoters in Recombinase Cacade