Objectif The human genome is largely composed of non-coding DNA, only 3-5% codes for exons. The majority of the non-coding DNA is however fundamental for the correct functioning and regulation of the genome. Transposable elements (TEs) like LINEs generated up to 50% of the human genome during evolution. They can mobilize, causing mutations but also conferring genomic plasticity. The generation of new insertions in the germ line led to the concept of TEs as ‘selfish’ DNA. However, inconsistent with their hereditary transmission, it was recently shown by the host lab and others that most of the action of TEs occurs in somatic cells during early embryogenesis.Thus, to deeply understand the impact of the somatic activity of LINE elements, we propose to develop and use an in vivo mouse model for LINE transposition. We will use human embryonic stem cells (hESCs) and tissue-specific iPSCs, containing an eGFP-marked LINE reporter cassette. Once injected into immuno-suppressed mice these cells will develop teratomas containing tissues of the 3 germ layers (endo, meso, ectoderm). For the proof of principle, the host lab has injected human embryonic carcinoma cells (hECs). With this model, we aim to answer the following: 1) Are LINEs active in all three germ layers?; 2) Are LINEs differentially regulated, depending on the germ layer?; 3) What’s the impact of LINE activity in the somatic tissues?To answer these questions, we will FACS-sort parts of the teratoma into ecto-, meso-, and endoderm-like cells and map the site of new LINE insertions during development, using deep sequencing. We aim to detect hot spots of LINE integration, allowing the analysis of possible genetic and phenotypic consequences. Imaging of the rest of the teratoma will further show us cell types where transposition occurs with higher frequency and possible phenotypic alterations. We therefore hope to detect new principles of genomic plasticity that could regulate gene expression. Champ scientifique natural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesdevelopmental biologymedical and health sciencesmedical biotechnologycells technologiesstem cellsnatural sciencesbiological sciencesgeneticsmutationnatural sciencesbiological sciencesgeneticsgenomes 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) FP7-PEOPLE-2011-IEF - Marie-Curie Action: "Intra-European fellowships for career development" Appel à propositions FP7-PEOPLE-2011-IEF Voir d’autres projets de cet appel Régime de financement MC-IEF - Intra-European Fellowships (IEF) Coordinateur FUNDACION PUBLICA ANDALUZA PROGRESO Y SALUD M.P. Contribution de l’UE € 168 896,40 Adresse AVENIDA AMERICO VESPUCIO 15 EDIF S2 41092 Sevilla Espagne Voir sur la carte Région Sur Andalucía Sevilla Type d’activité Research Organisations Contact administratif Esther Guirado Luna (Ms.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée