title: A riboswitch-based inducible gene expression system for mycobacteria.
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.
authors.affiliation: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Jessica C
authors.initials: JC
authors.lastname: Seeliger
authors.forename: Shana
authors.initials: S
authors.lastname: Topp
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Sogi
authors.forename: Mary L
authors.initials: ML
authors.lastname: Previti
authors.forename: Justin P
authors.initials: JP
authors.lastname: Gallivan
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D005434:Flow Cytometry; D015964:Gene Expression Regulation, Bacterial; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008264:Macrophages / Q000187:drug effects / Q000382:microbiology; D051379:Mice; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D011401:Promoter Regions, Genetic / Q000235:genetics; D058928:Riboswitch / Q000235:genetics*; D013806:Theophylline / Q000494:pharmacology; D014376:Tuberculosis / Q000382:microbiology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D058928:Riboswitch; D049452:Green Fluorescent Proteins; D013806:Theophylline; D002374:Catalase; C480353:katG protein, Mycobacterium tuberculosis; D001616:beta-Galactosidase