"Regenerative medicine using gene targeting by homologous recombination is one of the most powerful methods to achieve precise genome modifications. In the absence of double strand breaks the frequency of homologues recombination events at a target site is very low. A wide range of molecular tools are used to achieve targeting double strand breaks to increase the efficiency of homologues recombination. Meganucleases are a class of homing endonucleases that can recognize long DNA sequences with high specificity. This characteristic makes them one of the most promising tools to promote specific double strand breaks. Therefore the design of meganucleases able to recognize new DNA targets is emerging as a crucial area in the regenerative medicine. The huge effort of several labs has contributed to improve the designing process of new meganucleases. However, additonal efforts are urgently needed to understand the full potential of these proteins. The aim of this project is at first to deliver a more rational way to design meganucleases with new DNA specificity by incorporating in the designing stage DNA sequence-depended features that recently have been showed to influence protein binding, then deliver a library of engineered meganucleases able to recognize sequences of clinical interest and finally deliver new reagents to introduce such enzymes into iPS cells. All the findings will contribute to identify a potential new way to cure monogenic diseases and also build the bases to new therapeutic approach for multigenic disorder such as some types of leukemias and lymphomas. As a reintegration program and because of the collaborative nature of the proposal this action will support the candidate in returning to Europe and to take part in a new, vibrant and cutting-edge research field. The candidate will contribute to European research by using all the knowledge he has acquired during his experience in the USA."