Engineering a ligand-dependent RNA transcriptional activator. Export

@article{buskirk04, 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.}, address = {Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.}, author = {Buskirk, A. R. and Landrigan, A. and Liu, D. R.}, citeulike-article-id = {249109}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.chembiol.2004.05.017}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/15324817}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=15324817}, doi = {10.1016/j.chembiol.2004.05.017}, issn = {1074-5521}, journal = {Chem Biol}, keywords = {activator, aptamer, rna}, month = {August}, number = {8}, pages = {1157--1163}, posted-at = {2008-06-29 01:58:00}, priority = {0}, title = {Engineering a ligand-dependent RNA transcriptional activator.}, url = {http://dx.doi.org/10.1016/j.chembiol.2004.05.017}, volume = {11}, year = {2004} }

TY - JOUR ID - buskirk04 L3 - citeulike-article-id:249109 N2 - 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. IS - 8 JF - Chem Biol SN - 1074-5521 AD - Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA. EP - 1163 TI - Engineering a ligand-dependent RNA transcriptional activator. VL - 11 SP - 1157 KW - activator KW - aptamer KW - rna AU - Buskirk, AR AU - Landrigan, A AU - Liu, DR PY - 2004/08// UR - http://dx.doi.org/10.1016/j.chembiol.2004.05.017 ER -