A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila.
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.
Dietzl
Georg
G
Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria.
Chen
Doris
D
Schnorrer
Frank
F
Su
Kuan-Chung
KC
Barinova
Yulia
Y
Fellner
Michaela
M
Gasser
Beate
B
Kinsey
Kaolin
K
Oppel
Silvia
S
Scheiblauer
Susanne
S
Couto
Africa
A
Marra
Vincent
V
Keleman
Krystyna
K
Dickson
Barry J
BJ
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
D012333:RNA, Messenger; D043244:Ribonuclease III