Engineered riboregulators enable post-transcriptional control of gene expression

• Published in: Nature biotechnology Vol. 22; no. 7; pp. 841 - 847
• Main Authors: Collins, James J, Isaacs, Farren J, Dwyer, Daniel J, Ding, Chunming, Pervouchine, Dmitri D, Cantor, Charles R
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.07.2004; Nature Publishing Group
• Subjects: 5' Untranslated Regions - genetics; 5' Untranslated Regions - metabolism; Animals; Biological and medical sciences; Biotechnology; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Silencing; Genetic Engineering; Genetic technics; Green Fluorescent Proteins - analysis; Green Fluorescent Proteins - genetics; Methods. Procedures. Technologies; Modification of gene expression level; Molecular and cellular biology; Molecular genetics; Ribosomes - metabolism; RNA Processing, Post-Transcriptional; RNA, Messenger - analysis; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Untranslated - metabolism; Transcriptional Activation; Gene silencing; Regulation(control); RNA; Escherichia coli; Bacteria; Activation; Gene expression; Genetic translation; Enterobacteriaceae
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt986

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.