The present invention provides nucleic acid molecules, DNA constructs,
plasmids, and methods for post-transcriptional regulation of gene
expression using RNA molecules to both repress and activate translation
of an open reading frame. Repression of gene expression is achieved
through the presence of a regulatory nucleic acid element (the
cis-repressive RNA or crRNA) within the 5' untranslated region (5' UTR)
of an mRNA molecule. The nucleic acid element forms a hairpin (stem/loop)
structure through complementary base pairing. The hairpin blocks access
to the mRNA transcript by the ribosome, thereby preventing translation.
In particular, in embodiments of the invention designed to operate in
prokaryotic cells, the stem of the hairpin secondary structure sequesters
the ribosome binding site (RBS). In embodiments of the invention designed
to operate in eukaryotic cells, the stem of the hairpin is positioned
upstream of the start codon, anywhere within the 5' UTR of an mRNA. A
small RNA (trans-activating RNA, or taRNA), expressed in trans, interacts
with the crRNA and alters the hairpin structure. This alteration allows
the ribosome to gain access to the region of the transcript upstream of
the start codon, thereby activating transcription from its previously
repressed state.