A flow cytometry-based screen for synthetic riboswitches

• Type of Material: Article
• Authors: Sean A. Lynch, Emory University; Justin Gallivan, Emory University
• Language: English
• Date: 2008-11-25
• Publisher: Oxford University Press (OUP): Policy C - Option B
• Publication Version: Final Published Version
• Copyright Statement: © 2008 The Author(s)
• License: Creative Commons Attribution 2.0 Generic
• Final Published Version (URL): http://nar.oxfordjournals.org/content/37/1/184
• Title of Journal or Parent Work: Nucleic Acids Research
• ISSN: 0305-1048
• Volume: 37
• Issue: 1
• Start Page: 184
• End Page: 192
• Grant/Funding Information: J.P.G. is a Beckman Young Investigator, a Camille Dreyfus Teacher-Scholar, and an Alfred P. Sloan Research Fellow.; S.A.L. was supported by a National Science Foundation Problems and Research to Integrate Science and Mathematics (PRISM) Fellowship.; This work was supported by the National Institutes of Health (GM070740 to J.P.G.).; Funding for open access charge: National Institutes of Health (GM074070 to J.P.G.).
• Research Categories: Chemistry, General; Biology, Genetics

Abstract

Riboswitches regulate gene expression through direct, small molecule–mRNA interactions. The creation of new synthetic riboswitches from in vitro selected aptamers benefits from rapid, high-throughput methods for identifying switches capable of triggering dramatic changes in gene expression in the presence of a desired ligand. Here we present a flow cytometry-based screen for identifying synthetic riboswitches that induce robust increases in gene expression in the presence of theophylline. The performance characteristics of our newly identified riboswitches exceed those of previously described natural and synthetic riboswitches. Sequencing data and structure probing experiments reveal the ribosome binding site to be an important determinant of how well a switch performs and may provide insights into the design of new synthetic riboswitches.