title: A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila.
abstract: Forward genetic screens in model organisms have provided important insights into numerous aspects of development, physiology and pathology. With the availability of complete genome sequences and the introduction of RNA-mediated gene interference (RNAi), systematic reverse genetic screens are now also possible. Until now, such genome-wide RNAi screens have mostly been restricted to cultured cells and ubiquitous gene inactivation in Caenorhabditis elegans. This powerful approach has not yet been applied in a tissue-specific manner. Here we report the generation and validation of a genome-wide library of Drosophila melanogaster RNAi transgenes, enabling the conditional inactivation of gene function in specific tissues of the intact organism. Our RNAi transgenes consist of short gene fragments cloned as inverted repeats and expressed using the binary GAL4/UAS system. We generated 22,270 transgenic lines, covering 88% of the predicted protein-coding genes in the Drosophila genome. Molecular and phenotypic assays indicate that the majority of these transgenes are functional. Our transgenic RNAi library thus opens up the prospect of systematically analysing gene functions in any tissue and at any stage of the Drosophila lifespan.
authors.affiliation: Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria.
authors.forename: Georg
authors.initials: G
authors.lastname: Dietzl
authors.forename: Doris
authors.initials: D
authors.lastname: Chen
authors.forename: Frank
authors.initials: F
authors.lastname: Schnorrer
authors.forename: Kuan-Chung
authors.initials: KC
authors.lastname: Su
authors.forename: Yulia
authors.initials: Y
authors.lastname: Barinova
authors.forename: Michaela
authors.initials: M
authors.lastname: Fellner
authors.forename: Beate
authors.initials: B
authors.lastname: Gasser
authors.forename: Kaolin
authors.initials: K
authors.lastname: Kinsey
authors.forename: Silvia
authors.initials: S
authors.lastname: Oppel
authors.forename: Susanne
authors.initials: S
authors.lastname: Scheiblauer
authors.forename: Africa
authors.initials: A
authors.lastname: Couto
authors.forename: Vincent
authors.initials: V
authors.lastname: Marra
authors.forename: Krystyna
authors.initials: K
authors.lastname: Keleman
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Dickson
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D005260:Female; D015698:Genomic Library; D008297:Male; D009132:Muscles / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D009928:Organ Specificity; D034622:RNA Interference; D012333:RNA, Messenger; D043244:Ribonuclease III / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D043244:Ribonuclease III