Identifying genes that extend life span using a high-throughput screening system.
We developed a high-throughput functional genomic screening system that allows identification of genes prolonging lifespan in the baker's yeast Saccharomyces cerevisiae. The method is based on isolating yeast mother cells with a higher than average number of cell divisions as indicated by the number of bud scars on their surface. Fluorescently labeled wheat germ agglutinin (WGA) was used for specific staining of chitin, a major component of bud scars. The critical new steps in our bud-scar-sorting system are the use of small microbeads, which allows successive rounds of purification and regrowth of the mother cells (M-cell), and utilization of flow cytometry to sort and isolate cells with a longer lifespan based on the number of bud scars specifically labeled with WGA.
Chen
Cuiying
C
Department of Molecular Biomedical Research, Ghent University and Flanders Interuniversity Institute for Biotechnology (VIB), Ghent, Belgium.
Contreras
Roland
R
D000375:Aging / Q000235:genetics*; D003001:Cloning, Molecular; D005800:Genes, Fungal; D023281:Genomics; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*