An adaptor from translational to transcriptional control enables predictable assembly of complex regulation

Nat Methods. 2012 Nov;9(11):1088-94. doi: 10.1038/nmeth.2184. Epub 2012 Sep 30.

Abstract

Bacterial regulators of transcriptional elongation are versatile units for building custom genetic switches, as they control the expression of both coding and noncoding RNAs, act on multigene operons and can be predictably tethered into higher-order regulatory functions (a property called composability). Yet the less versatile bacterial regulators of translational initiation are substantially easier to engineer. To bypass this tradeoff, we have developed an adaptor that converts regulators of translational initiation into regulators of transcriptional elongation in Escherichia coli. We applied this adaptor to the construction of several transcriptional attenuators and activators, including a small molecule-triggered attenuator and a group of five mutually orthogonal riboregulators that we assembled into NOR gates of two, three or four RNA inputs. Continued application of our adaptor should produce large collections of transcriptional regulators whose inherent composability can facilitate the predictable engineering of complex synthetic circuits.

Publication types

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

MeSH terms

  • 5' Untranslated Regions / genetics
  • Base Sequence
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression Regulation, Bacterial*
  • Peptide Chain Initiation, Translational / physiology
  • Protein Sorting Signals / physiology
  • Synthetic Biology / methods
  • Transcription Elongation, Genetic / drug effects
  • Transcription Elongation, Genetic / physiology
  • Transcription, Genetic*

Substances

  • 5' Untranslated Regions
  • Protein Sorting Signals