Single-cell analysis of aneuploidy events using yeast whole chromosome painting probes (WCPPs)
| Autor: | Klaudia Kulak, Ewelina Golec, Jagoda Adamczyk, Anita Panek, Anna Deregowska, Anna Lewinska, Beata Miedziak, Maciej Wnuk |
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| Rok vydání: | 2015 |
| Předmět: |
Microbiology (medical)
Genetics education.field_of_study biology Population Saccharomyces cerevisiae Aneuploidy Cell cycle medicine.disease biology.organism_classification Microbiology Chromosome Painting Single-cell analysis Chromosome instability medicine Single-Cell Analysis Ploidy DNA Probes education Molecular Biology Cytometry In Situ Hybridization Fluorescence |
| Zdroj: | Journal of Microbiological Methods. 111:40-49 |
| ISSN: | 0167-7012 |
| DOI: | 10.1016/j.mimet.2015.01.022 |
| Popis: | Aneuploidy is considered a widespread genetic variation in such cell populations as yeast strains, cell lines and cancer cells, and spontaneous changes in the chromosomal copy number may have implications for data interpretation. Thus, aneuploidy monitoring is essential during routine laboratory practice, especially while conducting biochemical and/or gene expression analyses. In the present study, we constructed a panel of whole chromosome painting probes (WCPPs) to monitor aneuploidy in a single yeast Saccharomyces cerevisiae cell. The WCPP-based system was validated using "normal" haploid and diploid cells, as well as disomic cells both with and without cell synchronisation. FISH that utilised WCPPs was combined with DNA cell cycle analysis (imaging cytometry) to provide a detailed analysis of signal variability during the cell cycle. Chromosome painting can be utilised to detect spontaneously formed disomic chromosomes and study aneuploidy-promoting conditions. For example, the frequency of disomic chromosomes was increased in cells lacking NAD(+)-dependent histone deacetylase Sir2p compared with wild-type cells (p |
| Databáze: | OpenAIRE |
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