Functional characterization of Drosophila microRNAs by a novel in vivo library.
Animal microRNAs (miRNA) are implicated in the control of nearly all cellular functions. Due to high sequence redundancy within the miRNA gene pool, loss of most of these 21- to 24-bp long RNAs individually does not cause a phenotype. Thus, only very few miRNAs have been associated with clear functional roles. We constructed a transgenic UAS-miRNA library in Drosophila melanogaster that contains 180 fly miRNAs. This library circumvents the redundancy issues by facilitating the controlled misexpression of individual miRNAs and is a useful tool to complement loss-of-function approaches. Demonstrating the effectiveness of our library, 78 miRNAs induced clear phenotypes. Most of these miRNAs were previously unstudied. Furthermore, we present a simple system to create GFP sensors to monitor miRNA expression and test direct functional interactions in vivo. Finally, we focus on the miR-92 family and identify a direct target gene that is responsible for the specific wing phenotype induced by the misexpression of miR-92 family members.
Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
Claus
C
Schertel
Tobias
T
Rutishauser
Klaus
K
Förstemann
Konrad
K
Basler
D000818:Animals; D030801:Animals, Genetically Modified; D003001:Cloning, Molecular; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics*; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D015723:Gene Library; D049452:Green Fluorescent Proteins / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics; D010641:Phenotype; D014921:Wings, Animal / Q000502:physiology
D029721:Drosophila Proteins; C576274:MIRN92 microRNA, Drosophila; D035683:MicroRNAs; D008840:Microfilament Proteins; C540869:Sha protein, Drosophila; D049452:Green Fluorescent Proteins