Engineering precision RNA molecular switches

Engineering precision RNA molecular switches

Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520-8103

Edited by Larry Gold, NeXstar Pharmaceuticals, Inc., Boulder, CO, and approved January 15, 1999 (received for review September 29, 1998)

Abstract

Ligand-specific molecular switches composed of RNA were created by coupling preexisting catalytic and receptor domains via structural bridges. Binding of ligand to the receptor triggers a conformational change within the bridge, and this structural reorganization dictates the activity of the adjoining ribozyme. The modular nature of these tripartite constructs makes possible the rapid construction of precision RNA molecular switches that trigger only in the presence of their corresponding ligand. By using similar enzyme engineering strategies, new RNA switches can be made to operate as designer molecular sensors or as a new class of genetic control elements.

Footnotes

↵ * To whom reprint requests should be addressed. e-mail: ronald.breaker{at}yale.edu.
This paper was submitted directly (Track II) to the Proceedings Office.
↵ † The catalytic rate of an allosteric enzyme is regulated by the binding of an effector molecule to an allosteric binding site that is topographically distinct from the active site of the enzyme. Conformational changes brought about by the binding of a specific effector molecule result in modulation of the activity of the adjacent active site (18–21).
ABBREVIATION:

FMN,

flavin mononucleotide