An Updated Collection of Sequence Barcoded Temperature-Sensitive Alleles of Yeast Essential Genes
Open accessAbstract
Systematic analyses of essential gene function using mutant collections in Saccharomyces cerevisiae have been conducted using collections of heterozygous diploids, promoter shut-off alleles, through alleles with destabilized mRNA, destabilized protein, or bearing mutations that lead to a temperature-sensitive (ts) phenotype. We previously described a method for construction of barcoded ts alleles in a systematic fashion. Here we report the completion of this collection of alleles covering 600 essential yeast genes. This resource covers a larger gene repertoire than previous collections and provides a complementary set of strains suitable for single gene and genomic analyses. We use deep sequencing to characterize the amino acid changes leading to the ts phenotype in half of the alleles. We also use high-throughput approaches to describe the relative ts behavior of the alleles. Finally, we demonstrate the experimental usefulness of the collection in a high-content, functional genomic screen for ts alleles that increase spontaneous P-body formation. By increasing the number of alleles and improving the annotation, this ts collection will serve as a community resource for probing new aspects of biology for essential yeast genes.
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- Authors
- Kofoed, M., Milbury, K. L., Chiang, J. H., Sinha, S., Ben-Aroya, S., Giaever, G., Nislow, C., Hieter, P., & Stirling, P. C.
- Affiliations
- University of British Columbia (CA)
- BC Cancer Agency (CA)
- Faculty of Life Sciences Bar-Ilan University, Ramat-Gan, Israel
- Corresponding author
- Hieter, Philip
- Stirling, Peter C
- ORCID
- Venue
- G3 Genes|Genomes|Genetics
- Publisher
- Oxford University Press (OUP)
- Published
- 2015-09-01
- Volume
- 5
- Issue
- 9
- Pages
- 1879-1887
- ISSN
- 2160-1836 (ISSN-L)
- PMC
- PMC4555224
- OpenAlex
- W2214274340
- Cited by
- 48
- References
- 41
- Citations by year
- 2026: 3 · 2025: 8 · 2024: 6 · 2023: 5 · 2022: 3 · 2021: 5 · 2020: 9 · 2018: 2 · 2017: 2 · 2016: 4 · 2015: 1
- FWCI
- 1.30
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- G3 Genes Genomes Genetics (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Canadian Institutes of Health Research (MOP136982)
- National Cancer Institute (R01CA158162)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- temperature-sensitive allele, essential genes, Saccharomyces cerevisiae, mutant collections, mRNA destabilization, protein destabilization, amino acid substitutions, functional genomic screens, gene function
- MeSH terms
- Alleles, Diploidy, Models, Molecular, Mutation, Protein Conformation, Saccharomyces cerevisiae, Temperature, Yeasts, Genes, Essential, Genomics, Saccharomyces cerevisiae Proteins, DNA Barcoding, Taxonomic, High-Throughput Nucleotide Sequencing
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
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