Exploring the Dynamics and Mutational Landscape of Riboregulation with a Minimal Synthetic Circuit in Living Cells.
The regulation of gene expression, triggered by conformational changes in RNA molecules, is widely observed in cellular systems. Here, we examine this mode of control by means of a model-based design and construction of a fully synthetic riboregulatory device. We present a theoretical framework that rests on a simple energy model to predict the dynamic response of such a system. Following an equilibrium description, our framework integrates thermodynamic properties—anticipated with an RNA physicochemical model—with a detailed description of the intermolecular interaction. The theoretical calculations are confirmed with an experimental characterization of the action of the riboregulatory device within living cells. This illustrates, more broadly, the predictability of genetic robustness on synthetic systems, and the faculty to engineer gene expression programs from a minimal set of first principles.
Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Valencia, Spain; Institute of Systems and Synthetic Biology, Centre National de la Recherche Scientifique-Université d'Evry-Val d'Essonne, Évry, France. Electronic address: guillermo.rodrigo@csic.es.
Guillermo
G
Rodrigo
Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Valencia, Spain.
Eszter
E
Majer
School of Life Sciences, University of Warwick, Coventry, United Kingdom.
Satya
S
Prakash
Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Valencia, Spain.
José-Antonio
JA
Daròs
Institute of Systems and Synthetic Biology, Centre National de la Recherche Scientifique-Université d'Evry-Val d'Essonne, Évry, France; School of Life Sciences, University of Warwick, Coventry, United Kingdom.
Alfonso
A
Jaramillo
Centro Nacional Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Juan F
JF
Poyatos
D002470:Cell Survival; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics; D008957:Models, Genetic; D009154:Mutation; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics
D012329:RNA, Bacterial; D049452:Green Fluorescent Proteins; D012313:RNA