Figure 2.

Overview of humanization assays. For each human–yeast ortholog pair (curated from InParanoid and EggNOG), complementation assays in Saccharomyces cerevisiae were performed using three strategies: (i) genomic replacement at the native yeast loci via CRISPR-Cas9, (ii) temperature-sensitive inactivation of the yeast allele, and (iii) sporulation of a heterozygous diploid deletion strain followed by tetrad dissection. Complementing human orthologs were further characterized using various phenotypic assays, including quantitative growth measurements, environmental stress tests, mating, and segregation assays.