Construction of Caffeine-Inducible Gene Switches in Mammalian Cells

Methods Mol Biol. 2021;2312:159-168. doi: 10.1007/978-1-0716-1441-9_10.


Controlling gene expression in mammalian cells constitutes one of the core principles of mammalian synthetic biology. Especially for cell-based therapies, inducers of gene expression which demonstrate the highest degree of safety and patient adherence are needed. In this chapter, I describe methods to implement caffeine-controlled gene expression systems into mammalian cells. Using an array of different implementation strategies, from reconstituting transcription factors to activating endogenous signaling pathways, allows for a wide range of sensitivity and capacity of the resulting caffeine-responsive gene switches.

Keywords: Caffeine; Inducible expression; Receptors; Synthetic biology; Transcription factors.

MeSH terms

  • Caffeine / pharmacology*
  • Cell Culture Techniques
  • Cell Engineering*
  • Gene Expression Regulation / drug effects*
  • Genes, Switch*
  • HEK293 Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Signal Transduction
  • Synthetic Biology*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transfection


  • Intracellular Signaling Peptides and Proteins
  • Transcription Factors
  • Caffeine