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Author Notes:

Correspondence: Justin P. Gallivan, Email: justin.gallivan@emory.edu; Phone: 404-712-2171; Fax: 404-727-6586

Acknowledgments: We thank Pavan Bang for technical assistance, Professor Dale Edmondson for helpful discussions, and Drs. Doug Grubb and Wayne Patrick for critical readings of the manuscript.

DNA sequencing was performed at the NSF-supported Center for Fundamental and Applied Molecular Evolution (NSF-DBI-0320786).

Subjects:

Research Funding:

We are grateful to the Research Corporation, the Seaver Institute, the Arnold and Mabel Beckman Foundation, the NIH (R01-GM074070), and Emory University for financial support.

S.A.L. is an NSF-PRISM Fellow.

S.K.D. is grateful to Emory University for providing a George W. Woodruff Fellowship.

J.P.G. is a Beckman Young Investigator.

A High-Throughput Screen for Synthetic Riboswitches Reveals Mechanistic Insights into Their Function

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Journal Title:

Chemistry and Biology

Volume:

Volume 14, Number 2

Publisher:

, Pages 173-184

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Riboswitches are RNA-based genetic control elements that regulate gene expression in a ligand-dependent fashion without the need for proteins. The ability to create synthetic riboswitches that control gene expression in response to any desired small-molecule ligand will enable the development of sensitive genetic screens that can detect the presence of small molecules, as well as designer genetic control elements to conditionally modulate cellular behavior. Herein we present an automated high throughput screening method that identifies synthetic riboswitches that display extremely low background levels of gene expression in the absence of the desired ligand and robust increases in expression in its presence. Mechanistic studies reveal how these riboswitches function and suggest design principles for creating new synthetic riboswitches. We anticipate that the screening method and design principles will be generally useful for creating functional synthetic riboswitches.

Copyright information:

© 2007 Elsevier Ltd All rights reserved

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommerical-NoDerivs 3.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/3.0/).

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