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CRISPR/Cas9 Gene Targeting in Primary Mouse Bone Marrow-Derived Macrophages

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Cell Reprogramming for Immunotherapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2097))

Abstract

CRISPR-Cas9 technology allows for rapid, targeted genome editing at nearly any loci with limited off-target effects. Here, we describe a method for using retroviral transduction to deliver single-guide RNA to primary bone marrow-derived macrophages. This protocol allows for high-throughput reverse genetics assays in primary immune cells and is also compatible with retroviral systems for transgene expression.

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Author information

Authors and Affiliations

  1. Division for Protective Immunity, Department of Pathology and Laboratory Medicine, The Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    Will Bailis

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  1. Will Bailis
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Correspondence to Will Bailis .

Editor information

Editors and Affiliations

  1. Department of Pathology, Yale School of Medicine, New Haven, CT, USA

    Samuel G. Katz

  2. Department of Pathology, Yale School of Medicine, New Haven, CT, USA

    Peter M. Rabinovich

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Bailis, W. (2020). CRISPR/Cas9 Gene Targeting in Primary Mouse Bone Marrow-Derived Macrophages. In: Katz, S., Rabinovich, P. (eds) Cell Reprogramming for Immunotherapy. Methods in Molecular Biology, vol 2097. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0203-4_14

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  • DOI: https://doi.org/10.1007/978-1-0716-0203-4_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0202-7

  • Online ISBN: 978-1-0716-0203-4

  • eBook Packages: Springer Protocols

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