Reconstitution of human RNA interference in budding yeast.

Reconstitution of human RNA interference in budding yeast.
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DOI:
10.1093/nar/gkq1321
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发表时间:
2011-04
影响因子:
14.9
通讯作者:
Roth FP
Roth FP
中科院分区:
生物学2区
文献类型:
--
作者:
Suk K;Choi J;Suzuki Y;Ozturk SB;Mellor JC;Wong KH;MacKay JL;Gregory RI;Roth FP

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虽然RNA介导的干扰(RNAi)在真核生物中是一个广泛保守的过程,包括许多真菌,但在发芽酵母中却没有。三种人类蛋白,Ago2,Dester和TRBP,足以在体外重组RISC复合体。为了验证人RNAi基因的导入能否在酿酒酵母中重组RNAi,将编码这三种人类蛋白的基因导入酿酒酵母中。我们观察到siRNA和依赖于siRNA和RISC的靶基因GFP的沉默。因此,人的Ago2、DICER和TRBP可以在体内在酿酒酵母中功能性地重建人的RNAi,从而使得在一个简单的遗传模式生物中研究和使用人的RNAi途径成为可能。
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.
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