Sentinel interaction mapping - a generic approach for the functional analysis of human disease gene variants using yeast

• Published in: Disease models & mechanisms Vol. 13; no. 7
• Main Authors: Young, Barry P, Post, Kathryn L, Chao, Jesse T, Meili, Fabian, Haas, Kurt, Loewen, Christopher
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists Ltd 08.07.2020; The Company of Biologists
• Subjects: Arrays; Computational Biology; Databases, Genetic; Experiments; Gene Expression Regulation, Fungal; Genes; Genetic engineering; Genetic Predisposition to Disease; Genetic Variation; Genomics; Genotype & phenotype; High-Throughput Nucleotide Sequencing; human disease genes; Humans; Mutation; Phenotype; PTEN Phosphohydrolase - genetics; PTEN Phosphohydrolase - metabolism; Reproducibility of Results; Resource; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - growth & development; Saccharomyces cerevisiae - metabolism; Up-Regulation; variants; Yeast; Human disease genes; Variants; Yeast
• ISSN: 1754-8403; 1754-8411
• DOI: 10.1242/dmm.044560

Advances in sequencing technology have led to an explosion in the number of known genetic variants of human genes. A major challenge is to now determine which of these variants contribute to diseases as a result of their effect on gene function. Here, we describe a generic approach using the yeast to quickly develop gene-specific assays that can be used to quantify the level of function of a genetic variant. Using synthetic dosage lethality screening, 'sentinel' yeast strains are identified that are sensitive to overexpression of a human disease gene. Variants of the gene can then be functionalized in a high-throughput fashion through simple growth assays using solid or liquid media. Sentinel interaction mapping (SIM) has the potential to create functional assays for the large majority of human disease genes that do not have a yeast orthologue. Using the tumour suppressor gene as an example, we show that SIM assays can provide a fast and economical means to screen a large number of genetic variants.