Functional Genomics by comprehensive RNAi mutant fly bank in Drosophila

在本项目中，我们开发了诱导型RNAi系统，并将其应用于果蝇的14000个蛋白质编码基因。迄今为止，已构建了8300个基因对应的IR载体，建立了1.2万多个转化蝇系。为了验证这些RNAi苍蝇的有效性，并将其应用于研究基因的新功能，人们与世界各地的苍蝇研究人员进行了合作研究。这些合作涉及生命科学的许多学科，如细胞生长、细胞凋亡、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。通过这些合作出版的出版物多达20种，如下所示。通过合作，这些RNAi突变果蝇的具体特征也变得清晰。在胚胎阶段，诱导型RNAi对基因表达的抑制有时似乎弱于其他发育阶段，这可能是由于母体基因产物的数量巨大。在一些神经组织中，RNAi的表型也比用零突变镶嵌分析得到的表型弱。然而，在大多数情况下，RNAi果蝇可以特异性敲除目标基因，再现已知基因的典型表型，并揭示出功能未知的新基因表型。事实证明，筛选许多基因的预期表型是非常容易的，这意味着这些RNAi突变蝇库在基因功能分析的许多应用中是有用的。该项目培育的RNAi果蝇将于今年夏末在国立遗传研究所的库存中心向果蝇界开放。

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.