A comprehensive overview of yeast libraries and their role in advancing cell biology

Over the past two decades, genome-wide collections of mutants, or libraries, have revolutionized the fields of systems and cell biology by enabling systematic and high-throughput interrogation of gene and protein function. This has been especially prominent in the model yeast Saccharomyces cerevisiae. The unique genetic properties of yeast, combined with efficient genome engineering tools, have facilitated the creation of a large number of comprehensive collections of strains with targeted gene deletions, mutations, overexpressions, regulatable promoters, and protein tagging. These resources have enabled large-scale studies of cellular phenotypes, genetic and drug interactions, protein localization, protein–protein interactions, and much more. This review provides a comprehensive overview of the available systematic yeast libraries, highlighting their design, applications, and transformative impact on functional genomics. We detail how successive generations of libraries have addressed key challenges. Highlighting future applications, we discuss the potential integration of advanced fluorescent tools and machine learning approaches that promise to shape the next generation of libraries and establish yeast as a blueprint for systematic, dynamic, and predictive cell biology.