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Conditional Mutation of SMC5 in Mouse Embryonic Fibroblasts

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SMC Complexes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2004))

Abstract

The structural maintenance of chromosomes (SMC) complex, SMC5/6, is important for genome maintenance in all model eukaryotes. To date, the most extensive studies have focused on the roles of Smc5/6 in lower eukaryotes, such as yeast and fly. In the handful of studies that have used mammalian cells, siRNA was used by most to knockdown SMC5/6 components. RNAi methods have been very important for scientific progression, but they are hindered by incomplete silencing of protein expression and off-target effects. This chapter outlines the use of a conditional knockout approach in mouse embryonic fibroblasts to study the function of the SMC5/6 complex. These cell lines provide an alternative method to study the function and properties of the SMC5/6 complex in mammals.

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Acknowledgements

This work was supported by the National Institutes of Health (NIH) grant K99/R00 HD069458 to P.W.J.

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Author notes

  1. Himaja Gaddipati and Marina V. Pryzhkova contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA

    Himaja Gaddipati, Marina V. Pryzhkova & Philip W. Jordan

Authors
  1. Himaja Gaddipati
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  2. Marina V. Pryzhkova
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  3. Philip W. Jordan
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Corresponding author

Correspondence to Philip W. Jordan .

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Editors and Affiliations

  1. National Centre for Biological Sciences, Tata Institute of Fundamental Research (TIFR), Bangalore, India

    Anjana Badrinarayanan

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Gaddipati, H., Pryzhkova, M.V., Jordan, P.W. (2019). Conditional Mutation of SMC5 in Mouse Embryonic Fibroblasts. In: Badrinarayanan, A. (eds) SMC Complexes. Methods in Molecular Biology, vol 2004. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9520-2_4

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  • DOI: https://doi.org/10.1007/978-1-4939-9520-2_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9519-6

  • Online ISBN: 978-1-4939-9520-2

  • eBook Packages: Springer Protocols

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