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Functional Genomics by comprehensive RNAi mutant fly bank in Drosophila

果蝇综合 RNAi 突变体果蝇库的功能基因组学

基本信息

批准号：
15011253
负责人：
UEDA Ryu
金额：
$ 20.48万
依托单位：
National Institute of Genetics
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

In this project, we developed the 'inducible RNAi system and applied it to 14,000 protein-coding genes in Drosophila. To date, IR vectors corresponding 8,300 genes have been constructed and over 12.000 transformed fly lines have been established. To check the efficacy of these RNAi flies and to apply them to investigate new functions of the genes, collaborative researches have been carried out with fly researchers in the world. These collaborations include many subjects of life sciences, such as, cell growth, apoptosis, morphogenesis of cells and tissues, innate immunity, heart/trachea/alimentary canal development, neurogenesis, memory/learning, circadian rhythm, polyglutamin disease, Alzheimer disease, and glycobiology et etc. The publications through these collaborations are up to 20, which are listed below.The specific characters of these RNAi mutant flies have also become clear through the collaborations. In the embryonic stages, nock-down of gene expression by inducible RNAi seemed sometimes weaker than that at other developmental stages, probably because of the huge amount of the maternal gene product. Also in some neural tissues, the phenotype of the RNAi was weaker than that obtained by mosaic analysis using null mutation. However, in almost cases, the RNAi flies could knock-down the target gene specifically, reproduce a typical phenotype of the known gene, and reveal a new phenotype of the gene, whose function is not known. The fact that the screening of many genes for an anticipated phenotype proved to be very easy implies the usefulness of these RNAi mutant fly bank in many applications of gene function analyses. The RNAi flies produced in this project will be opened to the fly community till the end of this summer from the stock center in the National Institute of Genetics.

在这个项目中，我们开发了可诱导的RNAi系统，并将其应用于果蝇的14,000个蛋白质编码基因。到目前为止，已构建了与8,300个基因对应的IR载体，建立了12.000多个转化蝇系。为了检验这些RNAi苍蝇的有效性，并将它们应用于研究这些基因的新功能，世界各地的苍蝇研究人员开展了合作研究。这些合作涉及生命科学的许多学科，如细胞生长、细胞和组织的形态发生、先天免疫、心脏/气管/消化道发育、神经发生、记忆/学习、昼夜节律、多谷氨酰胺病、阿尔茨海默病和糖生物学等。在胚胎阶段，可诱导的RNAi对基因表达的抑制有时似乎比其他发育阶段弱，可能是由于母体基因产物的大量存在。此外，在一些神经组织中，RNAi的表型弱于使用零突变进行镶嵌分析所获得的表型。然而，在大多数情况下，RNAi果蝇可以特异性地敲除目标基因，复制已知基因的典型表型，并揭示该基因的新表型，其功能尚不清楚。事实证明，为一个预期的表型筛选许多基因是非常容易的，这表明这些RNAi突变体苍蝇库在基因功能分析的许多应用中是有用的。在这个项目中产生的RNAi苍蝇将在今年夏天底之前从国家遗传研究所的种群中心向苍蝇社区开放。