Synthetic gene expression perturbation systems with rapid, tunable, single-gene specificity in yeast

  • R. Scott Mcisaac
  • Benjamin L. Oakes
  • Xin Wang
  • Krysta A. Dummit
  • David Botstein
  • Marcus B. Noyes
Publication date
January 2013

Abstract

A general method for the dynamic control of single gene expression in eukaryotes, with no off-target effects, is a long-sought tool for molecular and systems biologists. We engineered two artificial transcription factors (ATFs) that contain Cys2His2 zinc-finger DNA-binding domains of either the mouse transcription factor Zif268 (9 bp of specifi-city) or a rationally designed array of four zinc fingers (12bp of specificity). These domains were expressed as fusions to the human estrogen receptor and VP16 activation domain. The ATFs can rapidly induce a single gene driven by a syn-thetic promoter in response to introduction of an otherwise inert hormone with no detectable off-target effects. In the absence of inducer, the synthetic promoter is...

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