ArticleRNA biology2013
Thermozymes: Synthetic RNA thermometers based on ribozyme activity.
Article in RNA biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Lipid membranes modulate the activity of RNA through sequence-dependent interactions.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Molecular simulations of the pistol ribozyme: unifying the interpretation of experimental data and establishing functional links with the hammerhead ribozyme.RNA (New York, N.Y.) · 2019Article
- RNA-DNA fibers and polygons with controlled immunorecognition activate RNAi, FRET and transcriptional regulation of NF-κB in human cells.Nucleic acids research · 2019Article
- Modular arrangement of regulatory RNA elements.RNA biology · 2017Review
- Programmable Genome Editing Tools and their Regulation for Efficient Genome Engineering.Computational and structural biotechnology journal · 2017Review
- Exploring the modular nature of riboswitches and RNA thermometers.Nucleic acids research · 2016Article
- RNAiFold2T: Constraint Programming design of thermo-IRES switches.Bioinformatics (Oxford, England) · 2016Article
- Fever as an important resource for infectious diseases research.Intractable & rare diseases research · 2016Review
- Thirty-five years of research into ribozymes and nucleic acid catalysis: where do we stand today?F1000Research · 2016Review
- Natural antisense RNAs as mRNA regulatory elements in bacteria: a review on function and applications.Cellular & molecular biology letters · 2016Review
- De novo design of heat-repressible RNA thermosensors in E. coli.Nucleic acids research · 2015Article
- Mechanistic insights into temperature-dependent regulation of the simple cyanobacterial hsp17 RNA thermometer at base-pair resolution.Nucleic acids research · 2015Article
- Complete RNA inverse folding: computational design of functional hammerhead ribozymes.Nucleic acids research · 2014Article
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Authors and funding
5 authors.
Funding
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Abstract
Synthetic biology approaches often combine natural building blocks to generate new cellular activities. Here, we make use of two RNA elements to design a regulatory device with novel functionality. The system is based on a hammerhead ribozyme (HHR) that cleaves itself to generate a liberated ribosome-binding site and, thus, permits expression of a downstream gene. We connected a temperature-responsive RNA hairpin to the HHR and, thus, generated a temperature-controlled ribozyme that we call thermozyme. Specifically, a Salmonella RNA thermometer (RNAT) known to modulate small heat shock gene expression by temperature-controlled base-pairing and melting was fused to the ribozyme. Following an in vivo screening approach, we isolated two functional thermozymes. In vivo expression studies and in vitro structure probing experiments support a mechanism in which rising temperatures melt the thermometer structure impairing the self-cleavage reaction of the ribozyme. Since RNA cleavage is necessary to liberate the RBS, these engineered thermozymes shut off gene expression in response to a temperature increase and, thus, act in a reverse manner as the natural RNAT. Our results clearly emphasize the highly modular nature and biotechnological potential of ribozyme-based RNA thermometers.
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