Antagonistic control of a dual-input mammalian gene switch by food additives

Nucleic Acids Res. 2014 Aug;42(14):e116. doi: 10.1093/nar/gku545. Epub 2014 Jul 16.

Abstract

Synthetic biology has significantly advanced the design of mammalian trigger-inducible transgene-control devices that are able to programme complex cellular behaviour. Fruit-based benzoate derivatives licensed as food additives, such as flavours (e.g. vanillate) and preservatives (e.g. benzoate), are a particularly attractive class of trigger compounds for orthogonal mammalian transgene control devices because of their innocuousness, physiological compatibility and simple oral administration. Capitalizing on the genetic componentry of the soil bacterium Comamonas testosteroni, which has evolved to catabolize a variety of aromatic compounds, we have designed different mammalian gene expression systems that could be induced and repressed by the food additives benzoate and vanillate. When implanting designer cells engineered for gene switch-driven expression of the human placental secreted alkaline phosphatase (SEAP) into mice, blood SEAP levels of treated animals directly correlated with a benzoate-enriched drinking programme. Additionally, the benzoate-/vanillate-responsive device was compatible with other transgene control systems and could be assembled into higher-order control networks providing expression dynamics reminiscent of a lap-timing stopwatch. Designer gene switches using licensed food additives as trigger compounds to achieve antagonistic dual-input expression profiles and provide novel control topologies and regulation dynamics may advance future gene- and cell-based therapies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Benzoic Acid / pharmacology
  • Cells, Cultured
  • Female
  • Food Additives / pharmacology*
  • Gene Expression Regulation / drug effects*
  • Gene Regulatory Networks / drug effects
  • Gene Silencing
  • HEK293 Cells
  • HeLa Cells
  • Herpes Simplex Virus Protein Vmw65 / genetics
  • Herpes Simplex Virus Protein Vmw65 / metabolism
  • Humans
  • Mice
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Synthetic Biology / methods
  • Transcriptional Activation
  • Transgenes
  • Vanillic Acid / pharmacology

Substances

  • Bacterial Proteins
  • CbaR protein, Comamonas testosteroni
  • Food Additives
  • Herpes Simplex Virus Protein Vmw65
  • Repressor Proteins
  • Benzoic Acid
  • Vanillic Acid