Isaacs Farren J, Dwyer Daniel J, Ding Chunming, Pervouchine Dmitri D, Cantor Charles R, Collins James J
Center for BioDynamics and Department of Biomedical Engineering, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA.
Nat Biotechnol. 2004 Jul;22(7):841-7. doi: 10.1038/nbt986. Epub 2004 Jun 20.
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.
最近的研究已经证明了RNA在细胞中重要的酶促、结构和调节作用。在此,我们展示了大肠杆菌中的一种转录后调控系统,该系统利用RNA来沉默和激活基因表达。我们在靶基因的核糖体结合位点正上游直接插入了一个互补顺式序列。转录时,这个顺式抑制序列会在mRNA的5'非翻译区形成一个茎环结构。该茎环结构会干扰核糖体结合,从而沉默基因表达。一种反式表达的小非编码RNA以高特异性靶向顺式抑制的RNA,导致茎环结构发生改变,从而激活表达。这种经过工程改造的核糖调节因子可能有助于深入了解基于RNA的内源性过程的作用机制,并可作为生物网络中可扩展的组件,能够与任何启动子或基因一起发挥作用,直接控制基因表达。