Engineering Genetic Control Mechanisms of Riboswitches

Yale University, New Haven CT

Investigators

Abstract

0323510 Ronald Breaker Yale University Engineering Genetic Control Mechanisms of Riboswitches Precision genetic control is an essential feature of living systems, as cells must respond to a multitude of biochemical signals and environmental cues by varying genetic expression patterns. Most known mechanisms of genetic control involve the use of protein factors that sense chemical or physical stimuli and then modulate gene expression by selectively interacting with the relevant DNA or messenger RNA sequence. Recently, it has been discovered that portions of certain mRNAs, called "riboswitches", serve as genetic regulation elements by forming direct RNA-metabolite complexes. This newly established form of genetic control is a critical mechanism by which biological information is accessed. Furthermore, the design of new RNA switches for use in diverse applications such as molecular sensing, designer genetic control and molecular computing could be enhanced by reverse engineering of natural riboswitches. A rigorous investigation of the biochemical properties of several metabolite-dependent riboswitches is being conducted in order to expand our understanding of the various mechanisms used by mRNAs to achieve metabolite recognition and genetic control. Several powerful biochemical techniques including an "in-line probing" assay is being used to give a detailed assessment of the structural changes that occur when ligands interact with allosteric RNAs. This will permit the development of a deeper understanding of how RNA acts to control genetic information. Furthermore, mutation analyses and genetic expression assays is being used to define the precise mechanisms by which ligand binding events are converted into control of genetic information expression. Finally, riboswitches are engineered to respond to other effector compounds that provide a means to more conveniently and thoroughly manipulate gene control in living systems. Such advances is providing enabling technologies for creating molecular computation systems for in vitro and in vivo uses, and are synergistic with ongoing efforts to create RNA logic gates.