Small interfering RNA-induced transcriptional gene silencing in human cells
Small interfering RNA-induced transcriptional gene silencing in human cells
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DOI：
10.1126/science.1101372
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发表时间：
2004-08-27
期刊：
影响因子：
56.9
通讯作者：
Looney, DJ
中科院分区：
文献类型：
--
作者：
Morris, KV;Chan, SWL;Looney, DJ
小干扰RNA （siRNA）和rnicroRNA在转录、转录后和/或翻译水平上沉默基因。利用人组织培养细胞，我们发现启动子导向的siRNA抑制了整合的前病毒延伸因子1 α (EF1A)启动子绿色荧光蛋白报告基因和内源性EF1A的转录。沉默与目标序列的DNA甲基化有关，它需要siRNA主动转运到细胞核中或通过慢病毒转导使核膜渗透。这些结果表明，sirna定向转录沉默在哺乳动物中是保守的，为抑制哺乳动物基因功能提供了一种手段。
Small interfering RNA (siRNA) and rnicroRNA silence genes at the transcriptional, posttranscriptional, and/or translational level. Using human tissue culture cells, we show that promoter-directed siRNA inhibits transcription of an integrated, proviral, elongation factor 1 alpha (EF1A) promoter-green fluorescent protein reporter gene and of endogenous EF1A. Silencing was associated with DNA methylation of the targeted sequence, and it required either active transport of siRNA into the nucleus or permeabilization of the nuclear envelope by lentiviral transduction. These results demonstrate that siRNA-directed transcriptional silencing is conserved in mammals, providing a means to inhibit mammalian gene function.