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Genetic Assays to Study Repeat Fragility in Saccharomyces cerevisiae

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Trinucleotide Repeats

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2056))

Abstract

Trinucleotide repeats are common in the human genome and can undergo changes in repeat number and cause length-dependent chromosome fragility. Expanded CAG repeats have been linked to over 14 human diseases and are considered hotspots for breakage and genomic rearrangement. Here we describe two Saccharomyces cerevisiae based assays that evaluate the rate of chromosome breakage that occurs within a repeat tract (fragility), with variations that allow the role of transcription to be evaluated. The first fragility assay utilizes end-loss and subsequent telomere addition as the main mode of repair of a yeast artificial chromosome (YAC). The second fragility assay relies on the fact that a chromosomal break stimulates recombination-mediated repair. A PCR-based assay can be used to evaluate instability of the repeat in the same conditions used to measure repeat fragility. These assays have contributed to understanding the genetic mechanisms that cause chromosome breaks and tract-length changes at unstable trinucleotide repeats.

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Acknowledgments

Work in our laboratory is currently supported by grants from the National Institute of Health (GM105473, GM122880) and National Science Foundation (MCB 1817499). We would like to thank past and present members of the C. H. Freudenreich and V.A. Zakian labs for their contributions to the development of these assays.

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Authors and Affiliations

  1. Department of Biology, Tufts University, Medford, MA, USA

    Erica J. Polleys & Catherine H. Freudenreich

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  1. Erica J. Polleys
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  2. Catherine H. Freudenreich
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Correspondence to Catherine H. Freudenreich .

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Editors and Affiliations

  1. Department Genomes & Genetics, Institut Pasteur, Paris, France

    Guy-Franck Richard

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Polleys, E.J., Freudenreich, C.H. (2020). Genetic Assays to Study Repeat Fragility in Saccharomyces cerevisiae. In: Richard, GF. (eds) Trinucleotide Repeats. Methods in Molecular Biology, vol 2056. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9784-8_5

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  • DOI: https://doi.org/10.1007/978-1-4939-9784-8_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9783-1

  • Online ISBN: 978-1-4939-9784-8

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