Patent ID: 9534224
Date: 2017-01-03
CPC Classifications: C12N,C12Q

Claim:
1. A recombinant prokaryotic cell comprising, a first exogenous mRNA nucleic acid sequence and a second exogenous mRNA nucleic acid sequence encoding a trans-activating RNA, wherein: a. the first exogenous mRNA nucleic acid sequence comprises a ribosome binding site (RBS) upstream of a nucleic acid sequence encoding a gene to be expressed, and a cis-repressive nucleic acid sequence located 5′ of the RBS, b. the second exogenous nucleic acid sequence comprises an inducible promoter operatively linked to a nucleic acid sequence encoding the trans-activating RNA, wherein the trans-activating RNA comprises (i) a 5′ non-stem nucleic acid sequence, comprising at least 4 nucleotides that are complementary to, or substantially complementary to at least 4 nucleotides of the cis-responsive nucleic acid sequence, and (ii) a stem-loop structure comprising a RNA duplex and loop; wherein: when the trans-activating RNA is not present, the cis-repressive nucleic acid sequence and RBS form the repressive stem-loop structure, wherein the repressive stem-loop structure blocks access of a ribosome to the RBS thereby repressing the translation of the gene, and when the trans-activating RNA is present, the 5′ non-stem nucleic acid sequence of the trans-activating RNA forms a duplex with at least 4 nucleotides of the cis-repressive nucleic acid sequence, wherein the duplex disrupts the repressive stem-loop structure to expose the RBS, thereby allowing translation of the gene.