Development of an artificial calcium-dependent transcription factor to detect sustained intracellular calcium elevation

ACS Synth Biol. 2014 Oct 17;3(10):717-22. doi: 10.1021/sb500070c. Epub 2014 Sep 9.

Abstract

The development of a synthetic transcription factor that responds to intracellular calcium signals enables analyzing cellular events at the single-cell level or "rewiring" the intracellular information networks. In this study, we developed the calcium-dependent transcription factor (CaTF), which was cleaved by calpain and then translocated to the nuclei where it induced reporter expression. Our results demonstrated that CaTF-mediated reporter expression was stable and responded to the intracellular calcium level and calpain activity. In addition, CaTF detected the sustained calcium increase that was induced by physiological stimulation with epidermal growth factor (EGF). These results suggest that CaTF could be a useful tool to analyze intracellular calcium signals and be an interface between an endogenous signal network and synthetic gene network.

Keywords: biosensor; cell therapy; interface; intracellular calcium; mammalian synthetic biology; synthetic transcription factor.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Calcium / metabolism*
  • Cell Line
  • Cricetinae
  • Epidermal Growth Factor / metabolism
  • Gene Expression
  • Genes, Reporter
  • Molecular Sequence Data
  • Protein Engineering
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Signal Transduction
  • Synthetic Biology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • Recombinant Proteins
  • Transcription Factors
  • Epidermal Growth Factor
  • Calcium