Modulation of gene expression by RNAi.

Modulation of gene expression by RNAi.
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RNAi 调节基因表达。

DOI:
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发表时间:
2005
期刊:
Methods in Molecular Medicine
影响因子:
--
通讯作者:
G. Demartino
G. Demartino
中科院分区:
--
文献类型:
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作者:
C. Wójcik;R. Fabunmi;G. Demartino

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RNA干扰(RNAi)是转录后基因沉默的一种形式,其中细胞内双链RNA(dsRNA)的存在导致具有互补序列的mRNA特异性降解。RNAi是一种自然现象,可以作为一种强有力的工具,通过在各种细胞类型中产生基因“敲低”来研究基因功能。RNAi由短干扰RNA(siRNA)介导,其在细胞内由长dsRNA产生。为了避免全身毒性作用,用通过化学合成或通过一系列酶促反应获得的21-23-bp长的siRNA直接转染哺乳动物细胞。本章涉及siRNA的设计,合成,转染,并在哺乳动物细胞培养系统的效率读出。的一般原理说明了功能性敲低p97/VCP(valosin-containing蛋白)在HeLa细胞中使用五种不同的siRNA序列。
RNA interference (RNAi) is a form of posttranscriptional gene silencing in which the presence within the cell of double-stranded RNA (dsRNA) leads to the specific degradation of mRNA with a complimentary sequence. RNAi is a natural phenomenon that can be exploited as a powerful tool to study gene function by generating gene "knockdowns" in various cell types. RNAi is mediated by short interfering RNAs (siRNAs), which are generated within cells from long dsRNAs. To avoid generalized toxic effects, mammalian cells are transfected directly with 21-23-bp-long siRNAs generated either by chemical synthesis or obtained by a series of enzymatic reactions. The present chapter deals with siRNA design, synthesis, transfection, and readout of efficiency in a mammalian cell culture system. The general principle is illustrated by the functional knockdown of p97/VCP (valosin-containing protein) in HeLa cells using five different siRNA sequences.
DOI: 10.1073/pnas.110149597
发表时间: 2000-06-06
影响因子: 11.1
作者:
Clemens, JC;Worby, CA;Dixon, JE
通讯作者: Dixon, JE
DOI: --
发表时间: 1979-10
期刊: The Journal of biological chemistry
影响因子: --
作者:
M. Minks;D. West;S. Benvin;C. Baglioni
通讯作者: M. Minks;D. West;S. Benvin;C. Baglioni