Engineered riboregulators enable post-transcriptional control of gene expression.
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.
Isaacs
Farren J
FJ
Center for BioDynamics and Department of Biomedical Engineering, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA.
Dwyer
Daniel J
DJ
Ding
Chunming
C
Pervouchine
Dmitri D
DD
Cantor
Charles R
CR
Collins
James J
JJ
D020121:5' Untranslated Regions / Q000235:genetics / Q000378:metabolism; D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D022661:RNA, Untranslated / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D015533:Transcriptional Activation
D020121:5' Untranslated Regions; D012333:RNA, Messenger; D022661:RNA, Untranslated; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins