Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
Nucleic Acids Res. 2011 Apr;39(7):e43. doi: 10.1093/nar/gkq1321. Epub 2011 Jan 19.
Although RNA-mediated interference (RNAi) is a widely conserved process among eukaryotes, including many fungi, it is absent from the budding yeast Saccharomyces cerevisiae. Three human proteins, Ago2, Dicer and TRBP, are sufficient for reconstituting the RISC complex in vitro. To examine whether the introduction of human RNAi genes can reconstitute RNAi in S. cerevisiae, genes encoding these three human proteins were introduced into S. cerevisiae. We observed both siRNA and siRNA- and RISC-dependent silencing of the target gene GFP. Thus, human Ago2, Dicer and TRBP can functionally reconstitute human RNAi in S. cerevisiae, in vivo, enabling the study and use of the human RNAi pathway in a facile genetic model organism.
尽管 RNA 介导的干扰（RNAi）是真核生物（包括许多真菌）中广泛保守的过程，但它不存在于出芽酵母酿酒酵母中。三种人类蛋白质，AGO2、Dicer 和 TRBP，足以在体外重新组成 RISC 复合物。为了研究引入人类 RNAi 基因是否可以在酿酒酵母中重新建立 RNAi，将编码这三种人类蛋白质的基因导入酿酒酵母。我们观察到 siRNA 和 siRNA 和 RISC 依赖性靶基因 GFP 的沉默。因此，人类 AGO2、Dicer 和 TRBP 可以在体内功能上重建人类 RNAi 在酿酒酵母中的功能，使人类 RNAi 途径的研究和应用在简单的遗传模式生物中成为可能。
