Investigating and modulating recombination control in Arabidopsis thaliana using synthetic biology

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Crossovers (COs) are the reciprocal strand exchange between non-sister homologous chromosomes during prophase I of meiosis. The frequency and distribution of COs is highly regulated with one CO being essential for fidelity in segregation at metaphase I. In many crop plants, the frequency and distribution of COs is often restricted. This is problematic for generating novel crop plants with advantageous alleles. In contrast Budding yeast has proportionally high levels of COs and does not show the same restrictions to CO distribution. Furthermore, CO formation is highly conserved between eukaryotes. In this project we will use synthetic biology to investigate and modulate meiotic recombination control in Arabidopsis thaliana. This approach will be used to investigate known meiotic genes and novel candidates. We will generate both chimeric Yeast-Arabidopsis protein constructs - exchanging Arabidopsis protein motifs or domains with yeast - and novel protein constructs from de-novo sequence assemblies. These constructs will be transformed into Arabidopsis and Yeast - using CRISPR/Cas9. We will analyse the products using state-of-the-art fluorescence microscopy (including confocal and super-resolution) as well as molecular marker techniques involving next generation sequencing and bioinformatics analysis. Through this synthetic biology system, we aim to investigate the function of meiotic regulatory proteins and modulate the imposition of COs. This work will test if CO control can be both artificially engineered and transferred across kingdom barriers. Through this we will further understanding of recombination control which will facilitate the development of novel tools for plant breeding.