Title: Systematic Gene-to-Phenotype Arrays: A High-Throughput Technique for Molecular Phenotyping.
Authors: Jaeger, Philipp A; Ornelas, Lilia; McElfresh, Cameron; Wong, Lily R; Hampton, Randolph Y; Ideker, Trey
Published In Mol Cell, (2018 01 18)
Abstract: We have developed a highly parallel strategy, systematic gene-to-phenotype arrays (SGPAs), to comprehensively map the genetic landscape driving molecular phenotypes of interest. By this approach, a complete yeast genetic mutant array is crossed with fluorescent reporters and imaged on membranes at high density and contrast. Importantly, SGPA enables quantification of phenotypes that are not readily detectable in ordinary genetic analysis of cell fitness. We benchmark SGPA by examining two fundamental biological phenotypes: first, we explore glucose repression, in which SGPA identifies a requirement for the Mediator complex and a role for the CDK8/kinase module in regulating transcription. Second, we examine selective protein quality control, in which SGPA identifies most known quality control factors along with U34 tRNA modification, which acts independently of proteasomal degradation to limit misfolded protein production. Integration of SGPA with other fluorescent readouts will enable genetic dissection of a wide range of biological pathways and conditions.
PubMed ID: 29351850
MeSH Terms: Cyclin-Dependent Kinase 8/genetics; Gene Regulatory Networks; Genotype; High-Throughput Nucleotide Sequencing/methods*; High-Throughput Screening Assays/methods*; Mediator Complex/genetics; Oligonucleotide Array Sequence Analysis; Phenotype; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae/genetics