RRC ID 54447
Author Li D, Roca M, Yuecel R, Lorenz A.
Title Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis.
Journal Chromosoma
Abstract Schizosaccharomyces pombe, also known as fission yeast, is an established model for studying chromosome biological processes. Over the years, research employing fission yeast has made important contributions to our knowledge about chromosome segregation during meiosis, as well as meiotic recombination and its regulation. Quantification of meiotic recombination frequency is not a straightforward undertaking, either requiring viable progeny for a genetic plating assay, or relying on laborious Southern blot analysis of recombination intermediates. Neither of these methods lends itself to high-throughput screens to identify novel meiotic factors. Here, we establish visual assays novel to Sz. pombe for characterizing chromosome segregation and meiotic recombination phenotypes. Genes expressing red, yellow, and/or cyan fluorophores from spore-autonomous promoters have been integrated into the fission yeast genomes, either close to the centromere of chromosome 1 to monitor chromosome segregation, or on the arm of chromosome 3 to form a genetic interval at which recombination frequency can be determined. The visual recombination assay allows straightforward and immediate assessment of the genetic outcome of a single meiosis by epi-fluorescence microscopy without requiring tetrad dissection. We also demonstrate that the recombination frequency analysis can be automatized by utilizing imaging flow cytometry to enable high-throughput screens. These assays have several advantages over traditional methods for analyzing meiotic phenotypes.
Volume 128(3)
Pages 385-396
Published 2019-9-1
DOI 10.1007/s00412-019-00691-y
PII 10.1007/s00412-019-00691-y
PMID 30739171
PMC PMC6823302
MeSH Chromosome Segregation* Chromosomes, Fungal* Flow Cytometry Fluorescent Antibody Technique Meiosis* Molecular Imaging Recombination, Genetic* Schizosaccharomyces / physiology* Spores, Fungal
IF 3.53
Times Cited 1
Yeast FY16936(yFS275) FY21620 FY16937(yFS286)