Figure 2. Artificial ZFs Can Be Used to Construct Synthetic Transcriptional Activators.
(A) Circuit design for synthetic transcriptional cascade. Synthetic transcription factors (sTFs) are expressed from an ATc-inducible GAL1 promoter (pGAL1). sTF activators are composed of artificial ZF arrays fused to a herpes simplex VP16 activation domain (AD) and a nuclear localization sequence (NLS). Upon induction, the sTF operates on a cognate synthetic promoter—minimal CYC1 promoter engineered with ZF binding sequences directly upstream of the TATA box—to direct the expression of a yeast-enhanced green fluorescent protein (yEGFP) reporter. Circuits were chromosomally integrated into S. cerevisiae.
(B) sTF activator circuits built from artificial ZF arrays activate transcription from cognate synthetic promoters in a dose-dependent fashion (ZF 37-12 shown here). Points represent mean values for three experiments ± SD. See also Figure S1.