A Riboswitch-Based Inducible Gene Expression System for Mycobacteria

• Type of Material: Article
• Authors: Jessica C. Seeliger, Stony Brook University; Shana Topp, University of California, Berkeley; Kimberly M. Sogi, University of California, Berkeley; Mary L. Previti, Stony Brook University; Justin Gallivan, Emory University; Carolyn R. Bertozzi, University of California, Berkeley
• Language: English
• Date: 2012-01-18
• Publisher: Public Library of Science
• Publication Version: Final Published Version
• Copyright Statement: © Seeliger et al.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1371/journal.pone.0029266
• Title of Journal or Parent Work: PLoS ONE
• ISSN: 1932-6203
• Volume: 7
• Issue: 1
• Start Page: e29266
• End Page: e29266
• Grant/Funding Information: ST was supported in part by a postdoctoral fellowship from the Cancer Research Institute (http://www.cancerresearch.org/).; This work was funded by National Institutes of Health (http://www.nih.gov/) grants AI51622 (to CRB) and GM074070 (to JPG).
• Research Categories: Biology, General; Chemistry, General

Abstract

Research on the human pathogen Mycobacterium tuberculosis (Mtb) would benefit from novel tools for regulated gene expression. Here we describe the characterization and application of a synthetic riboswitch-based system, which comprises a mycobacterial promoter for transcriptional control and a riboswitch for translational control. The system was used to induce and repress heterologous protein overexpression reversibly, to create a conditional gene knockdown, and to control gene expression in a macrophage infection model. Unlike existing systems for controlling gene expression in Mtb, the riboswitch does not require the co-expression of any accessory proteins: all of the regulatory machinery is encoded by a short DNA segment directly upstream of the target gene. The inducible riboswitch platform has the potential to be a powerful general strategy for creating customized gene regulation systems in Mtb.