Design of Synthetic Promoters for Gene Circuits in Mammalian Cells

Methods Mol Biol. 2017:1651:263-273. doi: 10.1007/978-1-4939-7223-4_19.

Abstract

Synthetic biology, the synthesis of engineering and biology, has rapidly matured and has dramatically increased the complexity of artificial gene circuits in recent years. The deployment of intricate synthetic gene circuits in mammalian cells requires the establishment of very precise and orthogonal control of transgene expression. In this chapter, we describe methods of modulating the expression of transgenes at the transcriptional level. Using cAMP-response element-binding protein (CREB)-dependent promoters as examples, a tool for the precise tuning of gene expression by using different core promoters and by varying the binding affinity of transcription factor operator sites is described.

Keywords: Cyclic AMP responsive elements; Inducible expression; Minimal promoters; Promoter design; Synthetic promoters.

MeSH terms

  • Animals
  • Cyclic AMP Response Element-Binding Protein / metabolism
  • Gene Regulatory Networks*
  • Genes, Reporter
  • HEK293 Cells
  • Humans
  • Promoter Regions, Genetic*
  • Signal Transduction
  • Synthetic Biology / methods
  • Transcriptional Activation*
  • Transfection / methods

Substances

  • Cyclic AMP Response Element-Binding Protein