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秀丽隐杆线虫卵巢中表达基因的RNA干扰分析。

RNAi analysis of genes expressed in the ovary of Caenorhabditis elegans.

作者信息

Piano F, Schetter A J, Mangone M, Stein L, Kemphues K J

机构信息

Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, USA.

出版信息

Curr Biol. 2000;10(24):1619-22. doi: 10.1016/s0960-9822(00)00869-1.

Abstract

As a step towards comprehensive functional analysis of genomes, systematic gene knockout projects have been initiated in several organisms [1]. In metazoans like C. elegans, however, maternal contribution can mask the effects of gene knockouts on embryogenesis. RNA interference (RNAi) provides an alternative rapid approach to obtain loss-of-function information that can also reveal embryonic roles for the genes targeted [2,3]. We have used RNAi to analyze a random set of ovarian transcripts and have identified 81 genes with essential roles in embryogenesis. Surprisingly, none of them maps on the X chromosome. Of these 81 genes, 68 showed defects before the eight-cell stage and could be grouped into ten phenotypic classes. To archive and distribute these data we have developed a database system directly linked to the C. elegans database (Wormbase). We conclude that screening cDNA libraries by RNAi is an efficient way of obtaining in vivo function for a large group of genes. Furthermore, this approach is directly applicable to other organisms sensitive to RNAi and whose genomes have not yet been sequenced.

摘要

作为迈向基因组全面功能分析的一步,已在多种生物中启动了系统的基因敲除项目[1]。然而,在像秀丽隐杆线虫这样的后生动物中,母体贡献可能会掩盖基因敲除对胚胎发生的影响。RNA干扰(RNAi)提供了一种替代性的快速方法来获取功能丧失信息,该信息也可以揭示所靶向基因的胚胎作用[2,3]。我们利用RNAi分析了一组随机的卵巢转录本,并鉴定出81个在胚胎发生中起关键作用的基因。令人惊讶的是,它们中没有一个位于X染色体上。在这81个基因中,68个在八细胞阶段之前就表现出缺陷,并且可以分为十个表型类别。为了存档和分发这些数据,我们开发了一个直接与秀丽隐杆线虫数据库(Wormbase)相连的数据库系统。我们得出结论,通过RNAi筛选cDNA文库是获得一大类基因体内功能的有效方法。此外,这种方法可直接应用于对RNAi敏感且基因组尚未测序的其他生物。

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