Phenotypic Activation to Discover Biological Pathways and Kinase Substrates

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Abstract
Observing the effects of gene perturbation on cells or organisms has long been a standard strategy in biological research. We developed a genome-wide gene overexpression library as a new tool for large-scale functional analysis in budding yeast. Previous large-scale genetic studies have focused on applications of the deletion mutant collection, which has arguably revolutionized the functional characterization of yeast genes. While extremely powerful, deletion mutant experiments are generally limited to the characterization of loss-of-function phenotypes. We have explored the potential for using the Synthetic Genetic Array (SGA) method, a platform for high-throughput genetic analysis, with a genome-wide “overexpression array”, in which each strain on the array overexpresses a unique yeast gene. The overexpression array enables gain-of-function phenotypes to be examined on a large scale, providing a unique insight into gene function and a novel source of reagents for the global mapping of genetic networks and functional relationships amongst genes and pathways. Understanding the molecular bases of overexpression phenotypes should also shed new light on the nature of genetic dominance.

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