Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
Nat Struct Mol Biol. 2012 Nov;19(11):1193-201. doi: 10.1038/nsmb.2392. Epub 2012 Sep 30.
We show that convergent transcription induces transcriptional gene silencing (TGS) in trans for both fission yeast and mammalian cells. This method has advantages over existing strategies to induce gene silencing. Previous studies in fission yeast have characterized TGS as a cis-specific process involving RNA interference that maintains heterochromatic regions such as centromeres. In contrast, in mammalian cells, gene silencing is known to occur through a post-transcriptional mechanism that uses exogenous short interfering RNAs or endogenous microRNAs to inactivate mRNA. We now show that the introduction of convergent transcription plasmids into either Schizosaccharomyces pombe or mammalian cells allows the production of double-stranded RNA from inserted gene fragments, resulting in TGS of endogenous genes. We predict that using convergent transcription to induce gene silencing will be a generally useful strategy and allow for a fuller molecular understanding of the biology of TGS.
我们表明,在有丝分裂酵母和哺乳动物细胞中, convergent transcription 可诱导转录基因沉默(TGS)。这种方法优于现有的诱导基因沉默的策略。先前在有丝分裂酵母中的研究已经将 TGS 描述为一种 cis 特异性过程,涉及 RNA 干扰,可维持着丝粒等异染色质区域。相比之下,在哺乳动物细胞中,基因沉默是通过一种转录后机制发生的,该机制使用外源性短干扰 RNA 或内源性 microRNAs 来使 mRNA 失活。我们现在表明,将 convergent transcription 质粒引入酿酒酵母或哺乳动物细胞中,可从插入的基因片段中产生双链 RNA,导致内源性基因的 TGS。我们预测,使用 convergent transcription 诱导基因沉默将是一种普遍有用的策略,并允许更全面地了解 TGS 的生物学。