A system for stable expression of short interfering RNAs in mammalian cells
A system for stable expression of short interfering RNAs in mammalian cells
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DOI：
10.1126/science.1068999
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发表时间：
2002-04-19
期刊：
影响因子：
56.9
通讯作者：
Agami, R
中科院分区：
文献类型：
--
作者：
Brummelkamp, TR;Bernards, R;Agami, R
由于缺乏有效产生稳定的功能丧失表型的方法，哺乳动物遗传学方法研究基因功能一直受到阻碍。我们在这里报告了一个新的载体系统，命名为pSUPER，它指导哺乳动物细胞中小干扰rna （sirna）的合成。我们发现，由该载体介导的siRNA表达导致基因表达的有效和特异性下调，导致目标基因的功能失活。使用该载体稳定表达sirna介导基因表达的持续抑制，从而允许分析在较长时间内发展的功能丧失表型。因此，pSUPER载体为分析多种哺乳动物细胞类型的基因功能提供了一个新的、强大的系统。
Mammalian genetic approaches to study gene function have been hampered by the lack of toots to generate stable loss-of-function phenotypes efficiently. We report here a new vector system, named pSUPER, which directs the synthesis of small interfering RNAs (siRNAs) in mammalian cells. We show that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes. Stable expression of siRNAs using this vector mediates persistent suppression of gene expression, allowing the analysis of loss-of-function phenotypes that develop over longer periods of time. Therefore, the pSUPER vector constitutes a new and powerful system to analyze gene function in a variety of mammalian cell types.