Boosting riboswitch efficiency by RNA amplification

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 natura...

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Bibliographic Details
Published in:Nucleic acids research 2015-05, Vol.43 (10), p.e66-e66
Main Authors: Emadpour, Masoumeh, Karcher, Daniel, Bock, Ralph
Format: Article
Language:English
Subjects:
DNA-Directed RNA Polymerases - genetics
Escherichia coli - genetics
Gene Expression Regulation
Genome, Chloroplast
Human immunodeficiency virus
Methods Online
nef Gene Products, Human Immunodeficiency Virus - genetics
nef Gene Products, Human Immunodeficiency Virus - metabolism
Nicotiana - genetics
Protein Biosynthesis
Riboswitch
Transgenes
Viral Proteins - genetics
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Summary: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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkv165