Developing tools for fission yeast quantitative genetics (360G-Wellcome-211682_Z_18_Z)

Complex traits play roles in various human diseases, with multiple genes being implicated, as identified by genome-wide association studies. The advent of genome-editing techniques such as CRISPR/Cas9 has ushered in the idea that these genes can be repaired to cure the disease, as shown for monogenic hereditary disorders. However, it is unclear whether these will be able to repair conditions influenced by multiple genetic factors, and the complexity of multicellular eukaryotes makes it difficult to predict the outcome. Herein, the fission yeast, Schizosaccharomyces pombe, could provide a useful tool, as its comparatively smaller genome and ability to breed strains facilitates studies of accurate quantitative genetics. However, this model requires improvements before it is suitable. The tendency for wild-type strains to self-mate causes difficulties when conducting inter-strain crosses. Therefore, we propose to engineer stable heterothallic strains to guarantee outbreeding using a counter-selectable Cre/loxP construct to remove the H1 box at the mat1 locus. We will then test to confirm that transformation has been successful and determine whether they are h+ or h-. This allows a simpler and more reliable approach to inter-strain mating, allowing for the production of larger library of genetically diverse strains for the analysis of complex traits.