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Genome Instability pp 613–629Cite as

Reporter-Based Synthetic Genetic Array Analysis: A Functional Genomics Approach for Investigating Transcript or Protein Abundance Using Fluorescent Proteins in Saccharomyces cerevisiae

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1672))

Abstract

Fluorescent reporter genes have long been used to quantify various cell features such as transcript and protein abundance. Here, we describe a method, reporter synthetic genetic array (R-SGA) analysis, which allows for the simultaneous quantification of any fluorescent protein readout in thousands of yeast strains using an automated pipeline. R-SGA combines a fluorescent reporter system with standard SGA analysis and can be used to examine any array-based strain collection available to the yeast community. This protocol describes the R-SGA methodology for screening different arrays of yeast mutants including the deletion collection, a collection of temperature-sensitive strains for the assessment of essential yeast genes and a collection of inducible overexpression strains. We also present an alternative pipeline for the analysis of R-SGA output strains using flow cytometry of cells in liquid culture. Data normalization for both pipelines is discussed.

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Authors and Affiliations

  1. The Donnelly Centre, University of Toronto, 160 College St., Toronto, ON, M5S 3E1, Canada

    Hendrikje Göttert, Mojca Mattiazzi Usaj, Adam P. Rosebrock & Brenda J. Andrews

  2. Department of Molecular Genetics, University of Toronto, 1 Kings College Circle, Toronto, ON, M5S 3E1, Canada

    Hendrikje Göttert, Adam P. Rosebrock & Brenda J. Andrews

  3. Department of Pathology and University Cancer Center, Stony Brook Medicine, Stony Brook, 11794, NY, USA

    Adam P. Rosebrock

Authors
  1. Hendrikje Göttert
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  2. Mojca Mattiazzi Usaj
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  3. Adam P. Rosebrock
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  4. Brenda J. Andrews
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Corresponding author

Correspondence to Brenda J. Andrews .

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Editors and Affiliations

  1. Dipartimento di Bioscienze, Università degli Studi di Milano, Milano, Milano, Italy

    Marco Muzi-Falconi

  2. Donnelly Centre and Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada

    Grant W Brown

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Göttert, H., Mattiazzi Usaj, M., Rosebrock, A.P., Andrews, B.J. (2018). Reporter-Based Synthetic Genetic Array Analysis: A Functional Genomics Approach for Investigating Transcript or Protein Abundance Using Fluorescent Proteins in Saccharomyces cerevisiae . In: Muzi-Falconi, M., Brown, G. (eds) Genome Instability. Methods in Molecular Biology, vol 1672. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7306-4_40

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  • DOI: https://doi.org/10.1007/978-1-4939-7306-4_40

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7305-7

  • Online ISBN: 978-1-4939-7306-4

  • eBook Packages: Springer Protocols

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