Roguev Assen, Ryan Colm J, Hartsuiker Edgar, Krogan Nevan J
Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94518
Systems Biology Ireland, University College Dublin, Belfield, Dublin 4, Ireland.
Cold Spring Harb Protoc. 2018 Feb 1;2018(2):pdb.top079905. doi: 10.1101/pdb.top079905.
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 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.
上位性作图是一种用于无偏研究基因功能的强大工具,它通过定量测量成对突变组合产生的表型来实现。当大规模进行时,这种方法已被用于为以前未表征的基因赋予功能、定义功能模块和途径,并研究它们之间的相互作用。这些实验严重依赖于通过系统生成一系列双突变体来快速采样突变等位基因二元组合的方法。现在,上位性作图技术存在于各种模型系统中。在这里,我们概述了不同的上位性作图技术,包括用于在裂殖酵母中收集定量遗传相互作用数据的粟酒裂殖酵母上位性作图器(PEM)系统。PEM系统包括一系列用于双突变体生成和表征的高通量选择步骤,它为广泛的生物学过程提供了见解,并促进了不同物种间遗传相互作用组的进化分析。