S133: Development of novel biosensors using bacterial riboswitches

RNA-based biosensors and regulatory devices have received significant attention for their potential in a broad array of synthetic biology and biotechnology applications. One of the difficulties in the creation of RNA sensors is linking the binding of an effector molecule to a readable output. In bacteria a widespread form of gene regulation called riboswitches performs this task with sufficient fidelity to control expression critical biosynthetic and transport proteins that are essential for normal cellular homeostasis.  We have developed a method for robust and easy mixing-and-matching of both natural and synthetic RNA receptors to biological regulatory domains that control transcription via an effector-dependent secondary structural switch.  This technique does not require the selection of device-specific communication modules necessary to transmit ligand binding to the regulatory domain allowing for rapid engineering of novel functional RNAs.  The resulting chimeras display robust regulatory activity using a simple in vitro transcription assay as well as regulatory activity in E. coli.