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Cold Spring Harb Protoc. 2018 Feb 1;2018(2). doi: 10.1101/pdb.top079905.

High-Throughput Quantitative Genetic Interaction Mapping in the Fission Yeast Schizosaccharomyces pombe.

Author information

1
Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94518 roguev@gmail.com nevan.krogan@ucsf.edu.
2
Systems Biology Ireland, University College Dublin, Belfield, Dublin 4, Ireland.
3
North West Cancer Research Institute, Bangor University, Bangor LL57 2UW, United Kingdom.

Abstract

Epistasis mapping, in which the phenotype that emerges from combining pairs of mutations is measured quantitatively, is a powerful tool for unbiased study of gene function. When performed at a large scale, this approach has been used to assign function to previously uncharacterized genes, define functional modules and pathways, and study their cross talk. These experiments rely heavily on methods for rapid sampling of binary combinations of mutant alleles by systematic generation of a series of double mutants. Epistasis mapping technologies now exist in various model systems. Here we provide an overview of different epistasis mapping technologies, including the pombe epistasis mapper (PEM) system designed for the collection of quantitative genetic interaction data in fission yeast Schizosaccharomyces pombe Comprising a series of high-throughput selection steps for generation and characterization of double mutants, the PEM system has provided insight into a wide range of biological processes as well as facilitated evolutionary analysis of genetic interactomes across different species.

PMID:
28733404
DOI:
10.1101/pdb.top079905

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