Engineering a ligand-dependent RNA transcriptional activator.
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Abstract | RNA has recently been shown to play diverse roles in gene regulation, including the small molecule-dependent inhibition of translation in prokaryotes. To create an artificial genetic switch that acts at the level of transcription, we fused a small molecule binding aptamer to a previously evolved RNA that activates transcription when localized to a promoter. We designed a conformational shift in which a helical element required for transcriptional activation was stabilized upon ligand binding. Selection and screening in S. cerevisiae optimized the linker region, generating an RNA that is 10-fold more active in the presence of tetramethylrosamine (TMR). TMR increases the activity of this evolved RNA in a graded, dose-dependent manner. Our results exemplify a strategy for controlling the activity of laboratory-evolved RNAs in living cells. |
Year of Publication | 2004
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Journal | Chem Biol
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Volume | 11
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Issue | 8
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Pages | 1157-63
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Date Published | 2004 Aug
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ISSN | 1074-5521
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DOI | 10.1016/j.chembiol.2004.05.017
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PubMed ID | 15324817
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