Engineering a ligand-dependent RNA transcriptional activator

Allen R. Buskirk, Angela Landrigan, David R. Liu

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)1157-1163
Number of pages7
JournalChemistry and Biology
Volume11
Issue number8
DOIs
StatePublished - Aug 2004
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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