Downward Julian
Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
Oncogene. 2004 Nov 1;23(51):8376-83. doi: 10.1038/sj.onc.1208073.
Over the past decade, 'RNA interference' has emerged as a natural mechanism of silencing of gene expression. This ancient cellular antiviral response can be manipulated to provide an effective research tool to knock down the level of expression of selected target genes, providing a very powerful new method for the analysis of cell signalling pathways. Systematic silencing of genes on a genome-wide scale using large rationally designed libraries targeting many thousands of genes provides a novel functional genomics approach to the investigation of many aspects of mammalian cell behaviour, including oncogenic transformation. Here, the different approaches taken to use RNA interference libraries to study the cancer phenotype will be considered, including both selective and high throughput screens and the use of both vector-based and synthetic oligonucleotide-based methods for inducing RNA interference. The advantages and drawbacks of the competing methodologies will be discussed. RNA interference library technology holds great promise for enabling somatic cell genetics in tissue culture systems. Whether it can provide significant new insights into cancer will be its greatest challenge.
在过去十年中，“RNA干扰”已成为一种使基因表达沉默的天然机制。这种古老的细胞抗病毒反应可被操控，以提供一种有效的研究工具来降低选定靶基因的表达水平，为细胞信号通路分析提供了一种非常强大的新方法。使用针对数千个基因的大型合理设计文库在全基因组范围内对基因进行系统性沉默，为研究哺乳动物细胞行为的许多方面，包括致癌转化，提供了一种新颖的功能基因组学方法。在此，将探讨利用RNA干扰文库研究癌症表型所采用的不同方法，包括选择性筛选和高通量筛选，以及使用基于载体和基于合成寡核苷酸的方法来诱导RNA干扰。还将讨论相互竞争的方法的优缺点。RNA干扰文库技术在组织培养系统中实现体细胞遗传学方面具有巨大潜力。它能否为癌症研究提供重大的新见解将是其最大的挑战。
