Toehold switches: de-novo-designed regulators of gene expression

Cell. 2014 Nov 6;159(4):925-39. doi: 10.1016/j.cell.2014.10.002. Epub 2014 Oct 23.

Abstract

Efforts to construct synthetic networks in living cells have been hindered by the limited number of regulatory components that provide wide dynamic range and low crosstalk. Here, we report a class of de-novo-designed prokaryotic riboregulators called toehold switches that activate gene expression in response to cognate RNAs with arbitrary sequences. Toehold switches provide a high level of orthogonality and can be forward engineered to provide average dynamic range above 400. We show that switches can be integrated into the genome to regulate endogenous genes and use them as sensors that respond to endogenous RNAs. We exploit the orthogonality of toehold switches to regulate 12 genes independently and to construct a genetic circuit that evaluates 4-input AND logic. Toehold switches, with their wide dynamic range, orthogonality, and programmability, represent a versatile and powerful platform for regulation of translation, offering diverse applications in molecular biology, synthetic biology, 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

  • Computer Simulation
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Gene Expression Regulation*
  • Gene Regulatory Networks*
  • RNA / chemistry*
  • Regulatory Sequences, Ribonucleic Acid
  • Synthetic Biology

Substances

  • Regulatory Sequences, Ribonucleic Acid
  • RNA