Nature Biotechnology
22, 841 - 847 (2004)
Published online: 20 June 2004; | doi:10.1038/nbt986
Engineered riboregulators enable post-transcriptional control of gene expressionFarren J Isaacs1, 2, 3, 4, Daniel J Dwyer1, 2, 5, Chunming Ding2, 3, Dmitri D Pervouchine1, 2, Charles R Cantor2, 3, 4
& James J Collins1, 2, 3, 41
Center for BioDynamics, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA. 2
Center for Advanced Biotechnology, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA. 3
Bioinformatics Program, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA. 4
Department of Biomedical Engineering, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA. 5
Department of Biology, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA.
Correspondence should be addressed to James J Collins jcollins@bu.eduRecent 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.
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