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Reengineering orthogonally selective riboswitches
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Dixon, N., Duncan, J. N., Geerlings, T., Dunstan, M. S., McCarthy, John E. G., Leys, D. and Micklefield, J. (2010) Reengineering orthogonally selective riboswitches. Proceedings of the National Academy of Sciences, Vol.107 (No.7). pp. 2830-2835. doi:10.1073/pnas.0911209107 ISSN 0027-8424.
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Official URL: http://dx.doi.org/10.1073/pnas.0911209107
Abstract
The ability to independently control the expression of multiple genes by addition of distinct small-molecule modulators has many applications from synthetic biology, functional genomics, pharmaceutical target validation, through to gene therapy. Riboswitches are relatively simple, small-molecule–dependent, protein-free, mRNA genetic switches that are attractive targets for reengineering in this context. Using a combination of chemical genetics and genetic selection, we have developed riboswitches that are selective for synthetic “nonnatural” small molecules and no longer respond to the natural intracellular ligands. The orthogonal selectivity of the riboswitches is also demonstrated in vitro using isothermal titration calorimetry and x-ray crystallography. The riboswitches allow highly responsive, dose-dependent, orthogonally selective, and dynamic control of gene expression in vivo. It is possible that this approach may be further developed to reengineer other natural riboswitches for application as small-molecule responsive genetic switches in both prokaryotes and eukaryotes.
| Item Type: | Journal Article | ||||
|---|---|---|---|---|---|
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
| Journal or Publication Title: | Proceedings of the National Academy of Sciences | ||||
| Publisher: | National Academy of Sciences | ||||
| ISSN: | 0027-8424 | ||||
| Official Date: | 2010 | ||||
| Dates: |
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| Volume: | Vol.107 | ||||
| Number: | No.7 | ||||
| Page Range: | pp. 2830-2835 | ||||
| DOI: | 10.1073/pnas.0911209107 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published |
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