Engineered riboregulators enable post-transcriptional control of gene expression

Nat Biotechnol. 2004 Jul;22(7):841-7. doi: 10.1038/nbt986. Epub 2004 Jun 20.

Abstract

Recent studies have demonstrated the important enzymatic, structural and regulatory roles of RNA in the cell. Here we present a post-transcriptional regulation system in Escherichia coli that uses RNA to both silence and activate gene expression. We inserted a complementary cis sequence directly upstream of the ribosome binding site in a target gene. Upon transcription, this cis-repressive sequence causes a stem-loop structure to form at the 5'-untranslated region of the mRNA. The stem-loop structure interferes with ribosome binding, silencing gene expression. A small noncoding RNA that is expressed in trans targets the cis-repressed RNA with high specificity, causing an alteration in the stem-loop structure that activates expression. Such engineered riboregulators may lend insight into mechanistic actions of endogenous RNA-based processes and could serve as scalable components of biological networks, able to function with any promoter or gene to directly control gene expression.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 5' Untranslated Regions / genetics
  • 5' Untranslated Regions / metabolism
  • Animals
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Silencing*
  • Genetic Engineering
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / genetics
  • RNA Processing, Post-Transcriptional*
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • RNA, Untranslated / metabolism*
  • Ribosomes / metabolism*
  • Transcriptional Activation*

Substances

  • 5' Untranslated Regions
  • RNA, Messenger
  • RNA, Untranslated
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins