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PMID: 12930946 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Developmental defects by antisense-mediated inactivation of micro-RNAs 2 and 13 in Drosophila and the identification of putative target genes.

Nucleic acids research ·Vol. 31 ·No. 17 ·2003-09-01 ·Pages 4973-80

Boutla A, Delidakis C, Tabler M

Abstract

Micro-RNAs are a class of small non-coding regulatory RNAs that impair translation by imperfect base pairing to mRNAs. For analysis of their cellular function we injected different miRNA-specific DNA antisense oligonucleotides in Drosophila embryos. In four cases we observed severe interference with normal development, one had a moderate impact and six oligonucleotides did not cause detectable phenotypes. We further used the miR-13a DNA antisense oligonucleotide as a PCR primer on a cDNA library template. In this experimental way we identified nine Drosophila genes, which are characterised by 3' untranslated region motifs that allow imperfect duplex formation with miR-13 or related miRNAs. These genes, which include Sos and Myd88, represent putative targets for miRNA regulation. Mutagenesis of the target motif of two genes followed by transfection in Drosophila Schneider 2 (S2) cells and subsequent reporter gene analysis confirmed the hypothesis that the binding potential of miR-13 is inversely correlated with gene expression.

MeSH Terms
Animals Base Sequence Binding Sites/genetics Cell Line DNA, Antisense/administration & dosage,genetics Drosophila/cytology,embryology,genetics Embryo, Nonmammalian/abnormalities,drug effects,metabolism Gene Expression Regulation, Developmental Genes, Insect/genetics Luciferases/genetics,metabolism MicroRNAs/genetics Mutation Oligonucleotides, Antisense/administration & dosage,genetics Protein Biosynthesis Recombinant Fusion Proteins/genetics,metabolism Transfection
Chemicals
DNA, Antisense MicroRNAs Oligonucleotides, Antisense Recombinant Fusion Proteins Luciferases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Boutla Alexandra
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Hellas, PO Box 1527, GR-71110 Heraklion/Crete, Greece.
Delidakis Christos
Tabler Martin
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkg707
Published
2003-09-01
Pages
4973-80
Language
English
Region
England
NLM ID
0411011
PMCID
PMC212806
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkg707 PMC Full Text View journal details
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