title: Boosting riboswitch efficiency by RNA amplification.
abstract: Riboswitches are RNA sensors that regulate gene expression in response to binding of small molecules. Although they conceptually represent simple on/off switches and, therefore, hold great promise for biotechnology and future synthetic biology applications, the induction of gene expression by natural riboswitches after ligand addition or removal is often only moderate and, consequently, the achievable expression levels are not very high. Here, we have designed an RNA amplification-based system that strongly improves the efficiency of riboswitches. We have successfully implemented the method in a biological system for which currently no efficient endogenous tools for inducible (trans)gene expression are available: the chloroplasts of higher plants. We further show that an HIV antigen whose constitutive expression from the chloroplast genome is deleterious to the plant can be inducibly expressed under the control of the RNA amplification-enhanced riboswitch (RAmpER) without causing a mutant phenotype, demonstrating the potential of the method for the production of proteins and metabolites that are toxic to the host cell.
authors.affiliation: Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476 Potsdam-Golm, Germany.
authors.forename: Masoumeh
authors.initials: M
authors.lastname: Emadpour
authors.affiliation: Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476 Potsdam-Golm, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Karcher
authors.affiliation: Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476 Potsdam-Golm, Germany rbock@mpimp-golm.mpg.de.
authors.forename: Ralph
authors.initials: R
authors.lastname: Bock
mesh_terms: D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D054628:Genome, Chloroplast; D014176:Protein Biosynthesis; D058928:Riboswitch; D014026:Nicotiana / Q000235:genetics; D019076:Transgenes; D014764:Viral Proteins / Q000235:genetics; D054311:nef Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism
chemical_list: D058928:Riboswitch; D014764:Viral Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; C514837:nef protein, Human immunodeficiency virus 1; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
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