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Mouse Cell Culture pp 77–95Cite as

Gene Editing of Mouse Embryonic and Epiblast Stem Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1940))

Abstract

Efficient and reliable methods for gene editing are critical for the generation of loss-of-gene function stem cells and genetically modified mice. Here, we outline the application of CRISPR-Cas9 technology for gene editing in mouse embryonic stem cells (mESCs) to generate knockout ESC chimeras for the fast-tracked analysis of gene function. Furthermore, we describe the application of gene editing directly to mouse epiblast stem cells (mEpiSCs) for modelling germ layer differentiation in vitro.

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Acknowledgments

Our work was supported by grants from the Australian Research Council (DP 160103651, DP 160100933), the National Health and Medical Research Council of Australia (1127976), and the late Mr. James Fairfax (Bridgestar Foundation). PPLT is a NHMRC Senior Principal Research Fellow (Grant 1110751).

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Authors and Affiliations

  1. The University of Sydney, Children’s Medical Research Institute, Westmead, NSW, Australia

    Tennille Sibbritt, Pierre Osteil, Xiaochen Fan, Jane Sun, Nazmus Salehin, Hilary Knowles, Joanne Shen & Patrick P. L. Tam

  2. The University of Sydney, School of Medical Sciences, Camperdown, NSW, Australia

    Tennille Sibbritt, Pierre Osteil & Patrick P. L. Tam

Authors
  1. Tennille Sibbritt
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  2. Pierre Osteil
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  3. Xiaochen Fan
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  4. Jane Sun
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  5. Nazmus Salehin
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  6. Hilary Knowles
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  7. Joanne Shen
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  8. Patrick P. L. Tam
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Corresponding author

Correspondence to Patrick P. L. Tam .

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Editors and Affiliations

  1. Lung Health Research Centre, Department of Pharmacology and Therapeutics, University of Melbourne, Melbourne, VIC, Australia

    Ivan Bertoncello

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Cite this protocol

Sibbritt, T. et al. (2019). Gene Editing of Mouse Embryonic and Epiblast Stem Cells. In: Bertoncello, I. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 1940. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9086-3_6

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  • DOI: https://doi.org/10.1007/978-1-4939-9086-3_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9085-6

  • Online ISBN: 978-1-4939-9086-3

  • eBook Packages: Springer Protocols

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