Project description DEENESFRITPL Delineating the process of DNA damage repair Genomic integrity is sustained through the DNA repair process, which corrects DNA damage. Failure to do so leads to ageing and cancer. The EU-funded RecPAIR project is focussing on the repair of double strand DNA breaks (DSBs), the most threatening DNA lesions. Scientists are interested to delineate the process of homologous recombination employed to repair DSBs using the intact DNA as a template. Using genome editing, they will mimic DSB formation, monitor chromosome dynamics and identify genes implicated in DSB repair. Collectively, the project will help elucidate one of the most fundamental processes in life, with important implications for health. Show the project objective Hide the project objective Objective The integrity of genetic information is central to life, yet the DNA is vulnerable to damage from internal and external sources. Incorrect repair of DNA damage drives mutagenesis, loss of genetic information, ageing, and cancer. Double strand DNA breaks (DSBs) are perhaps the most threatening DNA lesions, where the integrity of both strands of the DNA duplex is interrupted at the same position. In E. coli, faithful repair of DSBs is possible only through the homologous recombination (HR) pathway which uses replicated chromosome as a template to recover the information. At the center of HR lies an elusive search process, during which broken strand localises and pairs with the repair template.I will use a combination of CRISPR/dCas9 screening and in-situ genotyping of pooled library of strains to characterise the genetic landscape controlling the homology search. First, I will develop a low probability DSB induction method, to limit the DSB-formation to only a single chromosome per cell. Next, I will design and implement a whole-genome CRISPRi screen coupled to high-throughput sequencing and map the genes involved specifically in the homology directed repair of DSBs. The knowledge of the recombination-specific genes will allow to create a refined, high-quality phenotypic screen. In this screen the whole chromosome dynamics will be monitored and defects in the DNA movements will be characterised for each tested target with a microfluidic-based fluorescent microscopy. Each phenotype will be linked to a specific gene using the state-of-the-art in-situ phenotyping approach called DuMPLING. The functional characterisation of recombination genes will allow to conclude a molecular model of the search process in vivo. Fields of science natural sciencesbiological sciencesgeneticsDNAnatural sciencesphysical sciencesopticsmicroscopymedical and health sciencesclinical medicineoncologynatural sciencesbiological sciencesgeneticschromosomes Programme(s) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Topic(s) MSCA-IF-2018 - Individual Fellowships Call for proposal H2020-MSCA-IF-2018 See other projects for this call Funding Scheme MSCA-IF-EF-ST - Standard EF Coordinator UPPSALA UNIVERSITET Net EU contribution € 191 852,16 Address VON KRAEMERS ALLE 4 751 05 Uppsala Sweden See on map Region Östra Sverige Östra Mellansverige Uppsala län Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 191 852,16
