Objectif The most common form of editing in mammalian RNA is deamination of adenosine to inosine by ADAR1 and ADAR2. ADAR1p150, an interferon inducible adenosine deaminase, is also known to antagonize RNA interference (RNAi). To date, the mechanism by which this is carried out is unclear. For example, it is unknown if the embryonic lethality of ADAR1 knockout mice is related to antagonism of RNAi. Thus, I propose to study how ADAR1 antagonizes the RNAi pathway and if it interacts directly with RNAi components. Mutants of ADAR1 have been generated in the host laboratory. I propose to examine the activity of each mutant against RNAi in HEK 293 cells. These mutants will allow us to determine if RNA binding, localization or editing by ADAR1 is required for ADAR1 antagonism of RNAi. The effects on each branch of the RNAi pathway, siRNA and miRNA, will be elucidated using advanced techniques including miRNA detecting microArrays, cloning, dual-luciferase, and miRNA/siRNA reporters. Further, classical approaches of protein-protein interaction, and a recently optimized Tap-tagging approach, will be used to reveal if ADAR1 interacts directly with components of the RNAi machinery. This proposal supports collaboration between Dr. Bret Heale and Dr. Mary O’Connell. Dr. Heale received his PhD in June 2006 (USA) for his studies in the RNAi field, specifically related to siRNA and miRNA recognition of target sites, and in the course of his PhD published two first author publications (one in siRNA targeting and one in miRNA targeting) and authored a computer program to predict siRNA efficacy. Dr. Mary O’Connell (UK) has spent nearly twenty years working in the field of RNA editing, specifically examining ADARs. Thus, this proposal embodies the goals of the FP7 “People” work program to bring talented scientists to Europe, further European science, and to strengthen international ties. Champ scientifique natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomicsnatural sciencesbiological sciencesgeneticsRNA Mots‑clés Adenosine Deamination RNA editing RNA interference RNAi miRNA siRNA Programme(s) FP7-PEOPLE - Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Thème(s) PEOPLE-2007-4-2.IIF - Marie Curie Action: "International Incoming Fellowships" Appel à propositions FP7-PEOPLE-2007-4-2-IIF Voir d’autres projets de cet appel Régime de financement MC-IIF - International Incoming Fellowships (IIF) Coordinateur MEDICAL RESEARCH COUNCIL Contribution de l’UE € 169 957,94 Adresse 20 Park Crescent W1B 1AL LONDON Royaume-Uni Voir sur la carte Type d’activité Public bodies (excluding Research Organisations and Secondary or Higher Education Establishments) Contact administratif Mary O'connell (Dr.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée