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

Dis Model Mech. 2020 Jul 8;13(7):dmm044560. doi: 10.1242/dmm.044560.

Abstract

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 Saccharomyces cerevisiae to quickly develop gene-specific in vivo 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 PTEN as an example, we show that SIM assays can provide a fast and economical means to screen a large number of genetic variants.

Keywords: Human disease genes; Variants; Yeast.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Computational Biology
  • Databases, Genetic
  • Gene Expression Regulation, Fungal
  • Genetic Predisposition to Disease
  • Genetic Variation*
  • Genomics*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • PTEN Phosphohydrolase / genetics*
  • PTEN Phosphohydrolase / metabolism
  • Phenotype
  • Reproducibility of Results
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Up-Regulation

Substances

  • PTEN Phosphohydrolase
  • PTEN protein, human