Generation of a Fanconi anemia models in the pig by advanced genome engineering

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In the previous DFG-funded project Transposon-mediated transgenesis in the pig genome a highly efficient approach for germline transgenesis coupled with reliable transgene expression in the pig was established employing the SB transposon system. Here, we propose to establish an efficient in ovo genome engineering approach in the pig model by employing transposon, Cre/loxP and clustered regulated interspaced short palindromic repeat (CRISPR/Cas9) systems. The proposed experiments will exploit the transposon-transgenic animals established in the previous funding period to provide proof of principle for single step engineering of the pig genome with the aim to generate a pig disease model for Fanconi anemia. The available mouse models for Fanconi anemia do not recapitulate certain features of the disease. It is anticipated that a Fanconi pig will mirror the disease phenotype seen in humans and will become an important model for the development of curative therapies. The proposed project is based on previous own expertise in establishing transposon transgenesis, as well as generation of induced pluripotent stem cells in rodents and pigs.