TY - JOUR
T1 - High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe
AU - Roguev, Assen
AU - Wiren, Marianna
AU - Weissman, Jonathan S.
AU - Krogan, Nevan J.
N1 - Funding Information:
We thank A. Carr and O. Nielsen (pON177; University of Copenhagen) for providing reagents; S. Forsburg (FY1524 strain; University of Southern California) and K. Ekwall for reagents and discussion; C.J. Ingles, G. Cagney and D. Fiedler for critical reading of the manuscript and M. Shales for help with figures. J.S.W. is funded by the Howard Hughes Medical Institute, N.J.K. used funds from a Sandler Family Fellowship and A.R. was funded by a Howard Hughes Medical Institute postdoctoral fellowship. The work was also supported by funds from the California Institute of Quantitative Biology.
PY - 2007/10
Y1 - 2007/10
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/35848940244
U2 - 10.1038/nmeth1098
DO - 10.1038/nmeth1098
M3 - Article
C2 - 17893680
AN - SCOPUS:35848940244
SN - 1548-7091
VL - 4
SP - 861
EP - 866
JO - Nature Methods
JF - Nature Methods
IS - 10
ER -