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. Author manuscript; available in PMC: 2013 May 14.
Published in final edited form as: Cell. 2012 Aug 3;150(3):647–658. doi: 10.1016/j.cell.2012.05.045

Figure 2. Artificial ZFs Can Be Used to Construct Synthetic Transcriptional Activators.

Figure 2

(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.