Peroxisome Mini-Libraries: Systematic Approaches to Study Peroxisomes Made Easy

Methods Mol Biol. 2017:1595:305-318. doi: 10.1007/978-1-4939-6937-1_28.

Abstract

High-throughput methodologies have been extensively used in the budding yeast, Saccharomyces cerevisiae, to uncover fundamental principles of cell biology. Over the years, several collections of yeast strains (libraries) were built to enable systematic exploration of cellular functions. However, using these libraries experimentally is often labor intensive and restricted to laboratories that hold high throughput platforms. Utilizing the available full genome libraries we handpicked a subset of strains that represent all known and predicted peroxisomal proteins as well as proteins that have central roles in peroxisome biology. These smaller collections of strains, mini-libraries, can be rapidly and easily used for complicated screens by any lab. Since one of the libraries is built such that it can be easily modified in the tag, promoter and selection, we also discuss how these collections form the basis for creating a diversity of new peroxisomal libraries for future studies. Using manual tools, available in any yeast lab, coupled with few simple genetic approaches, we will show how these libraries can be "mixed and matched" to create tailor made libraries for screening. These yeast collections may now be exploited to study uncharted territories in the biology of peroxisomes by anyone, anywhere.

Keywords: Genetic screen; High content; Libraries; Peroxisome; SGA; SWAT; Saccharomyces cerevisiae; Yeast.

MeSH terms

  • Gene Library
  • Genetic Testing
  • High-Throughput Screening Assays
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Peroxisomes / metabolism*
  • Proteomics / methods
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Systems Biology* / methods

Substances

  • Membrane Proteins
  • Saccharomyces cerevisiae Proteins