Schizosaccharomyces pombe Methods and Protocols

This detailed volume explores techniques utilizing the fission yeast Schizosaccharomyces pombe, a powerful tractable model system for eukaryotic biology. The book begins with a section on microscopy and imaging, and it continues with chapters covering genome and plasmid editing, including the use of CRISPR/Cas9 technologies, as well as the analysis of DNA damage, repair, and recombination, and more. The volume also includes chapters with a focus on computational methods specific to the approaches. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Schizosaccharomyces pombe: Methods and Protocols serves as the ideal guide for researchers in the fission yeast community working with emerging technologies.

Dual Luciferase Reporter Assay in Schizosaccharomyces pombe.- High-Throughput Measurement of Single Fission Yeast Cell Volume Using Fluorescence Exclusion.- Measuring Cell Dimensions in Fission Yeast Using Machine Learning.- Ultrastructure Expansion Microscopy (U-ExM) of the Fission Yeast S. pombe.- Live Imaging of Fission Yeast Single Cell Lineages Using a Microfluidic Device.- Analysis of Cell Wall Mechanics in Fission Yeast.- Pil1 Co-Tethering Assay to Detect Protein-Protein Interactions in the Fission Yeast Schizosaccharomyces pombe.- Bi-Molecular Fluorescence Complementation as a Tool to Study Specific Dynamic Interactions Between Proteins in Fission Yeast.- Mapping Active RNA Polymerases in Proliferating and Quiescent Fission Yeast Cells Using Precision Run-On Sequencing.- Fast Fission Yeast Genome Editing by CRISPR/Cas9 Using Gap Repair and Fluoride Selection.- CRISPR–Cas9 Genome Editing in Auxotrophic and Non-Auxotrophic Fission Yeast Strains.- Proliferation Analyses of Conditional Knockdown Strains Using CRISPR Interference in Fission Yeast.- SIMPLE Method for Plasmid Editing.- Investigating Fission Yeast Mutagenesis Using Canavanine Sensitivity Assays.- A Rapidly Inducible DNA Double-Strand Break to Monitor Telomere Formation, DNA Repair, and Checkpoint Activation.- Targeted Forward Genetics: Saturating Mutational Analyses of Specific Target Loci Within the Genome.- Assessing Drug Sensitivity in Fission Yeast Using Half-Maximal Inhibitory Concentration (IC50) Assays.- ChIPmentation for Epigenomic Analysis in Fission Yeast.- Dissecting the Cell-Killing Mechanisms of Hydroxyurea Using Spot Assays.- Measuring Meiotic Recombination Frequency in Schizosaccharomyces pombe Using an Engineered Genetic Interval.- Label-Free Quantitative Phosphoproteomics in the Fission Yeast Schizosaccharomyces pombe.- Genome-Wide Profiling of Histone Modifications in Fission Yeast Using CUT&Tag.- Identifying Drug Sensitivities in Fission Yeast by Assessing Growth Kinetics.- An “In Schizo” Evaluation System to Screen for Human Kinesin-5 Inhibitors.- Automated Machine Learning Tools to Build Regression Models for Schizosaccharomyces pombe Omics Data.