Parallel profiling of mutants for chronological lifespan

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Strategic Research Priority: World Class Underpinning BioscienceUnderstanding the main players involved in cellular senescence is key to uncovering the mechanisms required for the promotion of healthy ageing. Fission yeast and budding yeast are distantly related organisms, both of which are exceptional model organisms used for ageing studies. The two yeasts therefore offer complementary insights into the ageing research. In this project, we employ barcode sequencing to S. pombe genome-wide deletion library of nonessential genes, accounting for more than 80% of the genome, in order to aid the discovery of mutants with long and short lifespans under different environmental conditions. We anticipate further utilization of these mutants for drug screen experiments. The barcodes are a unique 20-nucleotide sequence that laterally spans the KanMX4 cassette, which is inserted in the region of each of the deleted gene, and thus, the barcodes act as molecular marks with direct quantitation of the mutants during chronological assays. The mutant counts is captured by PCR and highlighted in the sequencing data extrapolated from DNA libraries. Methods that direct the project include, but not limited to, the setting and optimization of the barcode sequencing technique, DNA library preparation, fluorescence microscopy for cellular imaging along with FACS analysis.