Silencing the Drosophila ribosomal protein L14 gene using targeted RNA interference causes distinct somatic anomalies.
The Drosophila Minutes are haploinsufficient mutations that are defective in ribosomal protein (rp) production, resulting in short, thin bristles, delayed development and recessive lethality. In a Minute fly, the amount of rp gene messenger RNA (mRNA) is reduced to >or=50% of the normal amount of gene product, and becomes rate limiting for ribosome biogenesis, cell proliferation and growth. Haploinsufficiency increases the vulnerability to complete loss of gene function (homozygous null state) if hit by a second mutation. Because of the homozygous lethality, it has only been possible to study the effects of Minute mutations in heterozygous animals. To be able to study the consequences of a loss-of-function of an rp gene (0%>mRNA<50%) in developing and differentiated cells we used heritable RNA interference (RNAi) in combination with the yeast GAL4/UAS binary system to spatiotemporally knock down the ribosomal protein L14 (RpL14) gene. We show, at the RNA and phenotypic levels, that RNAi efficiently reduces RpL14 gene expression throughout development, causing lethality and distinct and dramatic somatic anomalies in both developing and differentiated cells.
Enerly
Espen
E
Division of Cell and Molecular Biology, Institute of Biology, University of Oslo, Blindernveien 31, N-0315 Oslo, Norway.
Larsson
Jan
J
Lambertsson
Andrew
A
D000015:Abnormalities, Multiple / Q000235:genetics*; D000818:Animals; D003433:Crosses, Genetic; D004330:Drosophila / Q000235:genetics* / Q000254:growth & development; D005123:Eye / Q000294:innervation / Q000378:metabolism / Q000648:ultrastructure; D005124:Eye Abnormalities / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005804:Genes, Lethal / Q000235:genetics; D008297:Male; D008855:Microscopy, Electron, Scanning; D009154:Mutation; D009421:Nervous System Malformations / Q000235:genetics; D010641:Phenotype; D034622:RNA Interference; D012269:Ribosomal Proteins / Q000235:genetics*
D012269:Ribosomal Proteins; C041174:ribosomal protein L14