Synergistic and Tunable Human Gene Activation by Combinations of Synthetic Transcription Factors

Nat Methods. 2013 Mar;10(3):239-42. doi: 10.1038/nmeth.2361. Epub 2013 Feb 3.

Abstract

Mammalian genes are regulated by the cooperative and synergistic actions of many transcription factors. In this study we recapitulate this complex regulation in human cells by targeting endogenous gene promoters, including regions of closed chromatin upstream of silenced genes, with combinations of engineered transcription activator-like effectors (TALEs). These combinations of TALE transcription factors induced substantial gene activation and allowed tuning of gene expression levels that will broadly enable synthetic biology, gene therapy and biotechnology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Binding Sites
  • Blotting, Western
  • Carcinoembryonic Antigen / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • GPI-Linked Proteins / genetics
  • Genetic Engineering / methods*
  • HEK293 Cells
  • Humans
  • Kallikreins / genetics*
  • Luciferases / genetics
  • Plasmids
  • Promoter Regions, Genetic
  • Prostate-Specific Antigen / genetics*
  • Real-Time Polymerase Chain Reaction
  • Receptor, ErbB-2 / genetics*
  • Transcription Factors / genetics*
  • Transcriptional Activation*
  • Transfection

Substances

  • CEACAM5 protein, human
  • Carcinoembryonic Antigen
  • GPI-Linked Proteins
  • Transcription Factors
  • Luciferases
  • ERBB2 protein, human
  • Receptor, ErbB-2
  • Kallikreins
  • kallikrein-related peptidase 3, human
  • Prostate-Specific Antigen