Full design automation of multi-state RNA devices to program gene expression using energy-based optimization

PLoS Comput Biol. 2013;9(8):e1003172. doi: 10.1371/journal.pcbi.1003172. Epub 2013 Aug 1.

Abstract

Small RNAs (sRNAs) can operate as regulatory agents to control protein expression by interaction with the 5' untranslated region of the mRNA. We have developed a physicochemical framework, relying on base pair interaction energies, to design multi-state sRNA devices by solving an optimization problem with an objective function accounting for the stability of the transition and final intermolecular states. Contrary to the analysis of the reaction kinetics of an ensemble of sRNAs, we solve the inverse problem of finding sequences satisfying targeted reactions. We show here that our objective function correlates well with measured riboregulatory activity of a set of mutants. This has enabled the application of the methodology for an extended design of RNA devices with specified behavior, assuming different molecular interaction models based on Watson-Crick interaction. We designed several YES, NOT, AND, and OR logic gates, including the design of combinatorial riboregulators. In sum, our de novo approach provides a new paradigm in synthetic biology to design molecular interaction mechanisms facilitating future high-throughput functional sRNA design.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Models, Genetic*
  • Protein Biosynthesis*
  • RNA / chemistry*
  • RNA / genetics*
  • RNA / metabolism
  • Reproducibility of Results
  • Ribosomes / metabolism
  • Synthetic Biology / methods*
  • Thermodynamics

Substances

  • RNA

Grants and funding

Work supported by the grants FP7-ICT-043338 (BACTOCOM) to AJ, and BIO2011-26741 (Ministerio de Economía y Competitividad, Spain) to JAD. GR is supported by an EMBO long-term fellowship co-funded by Marie Curie actions (ALTF-1177-2011), and TEL by a PhD fellowship from the AXA Research Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.