Nature Methods
- 4, 861 - 866 (2007)
Published online: 23 September 2007; | doi:10.1038/nmeth1098
High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombeAssen Roguev1, 2, Marianna Wiren1, 2, 3, Jonathan S Weissman1, 2, 4 & Nevan J Krogan1, 21
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 1700 4th Street San Francisco, California 94158, USA. 2
The California Institute for Quantitative Biomedical Research, University of California, San Francisco, 1700 4th Street San Francisco, California 94158, USA. 3
Karolinska Institutet, Department of Biosciences Novum, University College Sodertorn, Department of Natural Sciences, S-141 04 Huddinge, Sweden. 4
Howard Hughes Medical Institute, University of California, San Francisco, California 94158, USA.
Correspondence should be addressed to Nevan J Krogan krogan@cmp.ucsf.edu Epistasis analysis, which reports on the extent to which the function of one gene depends on the presence of a second, is a powerful tool for studying the functional organization of the cell. Systematic genome-wide studies of epistasis, however, have been limited, with the majority of data being collected in the budding yeast, Saccharomyces cerevisiae. Here we present two 'pombe epistasis mapper' strategies, PEM-1 and PEM-2, which allow for high-throughput double mutant generation in the fission yeast, S. pombe. These approaches take advantage of a previously undescribed, recessive, cycloheximide-resistance mutation. Both systems can be used for genome-wide screens or for the generation of high-density, quantitative epistatic miniarray profiles (E-MAPs). Since S. cerevisiae and S. pombe are evolutionary distant, this methodology will provide insight into conserved biological pathways that are present in S. pombe, but not S. cerevisiae, and will enable a comprehensive analysis of the conservation of genetic interaction networks.
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