This doctoral research study funded by the SNF (Swiss National Science Foundation) comprises a comparative approach and includes bovine and roe deer as model species. We will characterize the molecular patterns during embryo development, translate previously described dormancy markers established in somatic cells to embryonic stem cells as a screening system, and elucidate the impact of dormancy inducing factors on cell cycle progression. We will then introduce CRISPRi in ESCs to screen for novel dormancy inducing factors. Novel chemical inhibitors targeting the discovered factors are applied to reversibly inhibit the identified pathways in bovine stem cells and embryos produced in vitro. Next-generation transcriptome and methylome sequencing will critically assess the developmental competence of the embryos after reversible developmental arrest. Identifying molecular markers of cellular dormancy will allow the development of culture conditions under which embryos may be kept in a reversible diapause-like stage. It may additionally provide implications for further livestock and endangered wildlife species, as well as the field of stem cell research and human medicine.
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