A strategy to analyze the phenotypic consequences of inhibiting the association of an RNA-binding protein with a specific RNA.: Targeted inhibition of RNA-protein interaction
| العنوان: | A strategy to analyze the phenotypic consequences of inhibiting the association of an RNA-binding protein with a specific RNA.: Targeted inhibition of RNA-protein interaction |
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| المؤلفون: | Luc Paillard, Marie Cibois, Audrey Vallée, Carole Gautier-Courteille |
| المساهمون: | Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Université européenne de Bretagne - European University of Brittany (UEB) |
| المصدر: | RNA RNA, Cold Spring Harbor Laboratory Press, 2010, 16 (1), pp.10-5. ⟨10.1261/rna.1742610⟩ |
| بيانات النشر: | Cold Spring Harbor Laboratory, 2009. |
| سنة النشر: | 2009 |
| مصطلحات موضوعية: | Embryo, Nonmammalian, Xenopus, Embryonic Development, RNA-binding protein, Xenopus Proteins, MESH: Base Sequence, MESH: Gene Targeting, MESH: Phenotype, Models, Biological, Xenopus laevis, 03 medical and health sciences, MESH: Oligonucleotides, Antisense, MESH: Xenopus laevis, MESH: Gene Expression Regulation, Developmental, Animals, MESH: Protein Binding, MESH: Embryonic Development, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, RNA, Messenger, Binding site, Letter to the Editor, MESH: Xenopus Proteins, Molecular Biology, CELF1 Protein, Derepression, MESH: RNA, Messenger, 030304 developmental biology, 0303 health sciences, Gene knockdown, Messenger RNA, Base Sequence, biology, 030302 biochemistry & molecular biology, MESH: Models, Biological, Gene Expression Regulation, Developmental, RNA-Binding Proteins, MESH: Embryo, Nonmammalian, RNA, Oligonucleotides, Antisense, biology.organism_classification, MESH: Gene Knockdown Techniques, Molecular biology, Hairless, Phenotype, MESH: RNA-Binding Proteins, Gene Knockdown Techniques, Gene Targeting, Protein Binding |
| الوصف: | International audience; Targeted inactivations of RNA-binding proteins (RNA-BPs) can lead to huge phenotypical defects. These defects are due to the deregulation of certain mRNAs. However, we generally do not know, among the hundreds of mRNAs that are normally controlled by one RNA-BP, which are responsible for the observed phenotypes. Here, we designed an antisense oligonucleotide ("target protector") that masks the binding site of the RNA-BP CUG-binding protein 1 (CUGBP1) on the mRNA Suppressor of Hairless [Su(H)] that encodes a key player of Notch signaling. We showed that injecting this oligonucleotide into Xenopus embryos specifically inhibited the binding of CUGBP1 to the mRNA. This caused the derepression of Su(H) mRNA, the overexpression of Su(H) protein, and a phenotypic defect, loss of somitic segmentation, similar to that caused by a knockdown of CUGBP1. To demonstrate a causal relationship between Su(H) derepression and the segmentation defects, a rescue experiment was designed. Embryonic development was restored when the translation of Su(H) mRNA was re-repressed and the level of Su(H) protein was reduced to a normal level. This "target protector and rescue assay" demonstrates that the phenotypic defects associated with CUGBP1 inactivation in Xenopus are essentially due to the deregulation of Su(H) mRNA. Similar approaches may be largely used to uncover the links between the phenotype caused by the inactivation of an RNA-BP and the identity of the RNAs associated with that protein. |
| تدمد: | 1469-9001 1355-8382 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5d47a816081fbabfa8adc23acced1162Test https://doi.org/10.1261/rna.1742610Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....5d47a816081fbabfa8adc23acced1162 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14699001 13558382 |
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