Engineering a ligand-dependent RNA transcriptional activator.
Chem Biol
; 11(8): 1157-63, 2004 Aug.
Article
em En
| MEDLINE
| ID: mdl-15324817
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.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
RNA
/
Engenharia Genética
/
Ativação Transcricional
Idioma:
En
Ano de publicação:
2004
Tipo de documento:
Article