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Use of Gene Targeting to Study Recombination in Mammalian Cell DNA Repair Mutants

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DNA Repair Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 920))

Abstract

The study of gene function has been greatly facilitated by the development of strategies to modify genomic DNA. Gene targeting is one of the most successfully applied techniques used to examine the roles of specific genes in a wide variety of model systems from yeast to mammals. Our laboratory has pioneered the use of the Chinese hamster ovary (CHO) cell culture model system to study pathways of DNA repair and recombination at the hemizygous CHO APRT locus. By using a simple and effective gene targeting method, we have generated a number of DNA repair-deficient cell lines that have been used in targeted recombination experiments to investigate pathways of recombinational repair in somatic mammalian cells. These methods can be readily customized to generate a variety of cell lines deficient in specific genes of interest and can be applied to study the roles of other DNA repair proteins in pathways of mammalian recombinational repair.

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Acknowledgements

We thank Becky Brooks for assistance in manuscript preparation. This work was supported by PHS grant CA097175 from the National Cancer Institute.

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Authors and Affiliations

  1. Department of Molecular Carcinogenesis, Medical University of South Carolina, Charleston, SC, USA

    Jennifer J. Rahn

  2. Department of Molecular Carcinogenesis, University of Texas M.D. Anderson Cancer Center, Smithville, TX, USA

    Gerald M. Adair & Rodney S. Nairn

Authors
  1. Jennifer J. Rahn
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  2. Gerald M. Adair
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  3. Rodney S. Nairn
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Corresponding author

Correspondence to Rodney S. Nairn .

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Editors and Affiliations

  1. , Department of Molecular Biology, Aarhus University, C.F Møllers Allé bygning 1130, Århus C, 8000, Denmark

    Lotte Bjergbæk

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Rahn, J.J., Adair, G.M., Nairn, R.S. (2012). Use of Gene Targeting to Study Recombination in Mammalian Cell DNA Repair Mutants. In: Bjergbæk, L. (eds) DNA Repair Protocols. Methods in Molecular Biology, vol 920. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-998-3_31

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  • DOI: https://doi.org/10.1007/978-1-61779-998-3_31

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-997-6

  • Online ISBN: 978-1-61779-998-3

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