Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Highlight
  • Published:

Genetic screens

Epistasis on the double

Nature Reviews Genetics volume 8, pages 826–827 (2007)Cite this article

The protocol simply involves crossing two haploid strains of yeast, each containing a different mutation, and then selecting for haploid progeny that carry two mutations. The initial cross is standard: it yields diploid cells that, after meiosis and sporulation, yield haploid progeny (see figure). Then comes the tricky part: singling out the double-mutant haploid cells. This requires three selection steps: one to remove any remaining diploid cells that have not undergone meiosis (as these would outgrow the double-mutant haploid cells); one to remove one or the other mating type, to avoid renewed mating between cells; and finally one to remove wild-type and single-mutant strains.

The last of these selection systems is straightforward as it involves using off-the-shelf antibiotic-resistance markers, which are linked to the mutations in the starting strains. The markers for the first two selection steps, however, had to be custom-made. To select against diploids, the authors induced a recessive mutation in a ribosomal protein that confers resistance to cycloheximide, which would be added to the medium to remove any heterozygous diploid cells. Selecting against one of the two mating strains was done either by placing an auxotrophic marker under the control of a promoter for a mating-type-specific pheromone or by introducing a selectable marker near the mating-type locus itself.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Epistasis on the double.

References

ORIGINAL RESEARCH PAPER

  1. Roguev, A. et al. High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe. Nature Methods 23 September 2007 (doi: 10.1038/nmeth1098).

    Article  CAS  PubMed  Google Scholar 

FURTHER READING

  1. Forsburg, S. L. The art and design of genetic screens: yeast. Nature Rev. Genet. 2, 659–668 (2001)

    Article  CAS  PubMed  Google Scholar 

  2. Collins, S. R. et al. Functional dissection of protein complexes involved in yeast chromosome biology using a genetic interaction map. Nature 446, 806–810 (2007)

    Article  CAS  PubMed  Google Scholar 

Download references

Authors
  1. Tanita Casci
    View author publications

    You can also search for this author in PubMed Google Scholar

Related links

Related links

WEB SITE

Nevan Krogan's Laboratory

Rights and permissions

Reprints and permissions

About this article

Cite this article

Casci, T. Epistasis on the double. Nat Rev Genet 8, 826–827 (2007). https://doi.org/10.1038/nrg2221

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nrg2221

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing