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Protocols for CRISPR-Cas9 Screening in Lymphoma Cell Lines

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Lymphoma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1956))

Abstract

Genome-wide screens are a powerful technique to dissect the complex network of genes regulating diverse cellular phenotypes. The recent adaptation of the CRISPR-Cas9 system for genome engineering has revolutionized functional genomic screening. Here, we present protocols used to introduce Cas9 into human lymphoma cell lines, produce high-titer lentivirus of a genome-wide sgRNA library, transduce and culture cells during the screen, isolate genomic DNA, and prepare a custom library for next-generation sequencing. These protocols were tailored for loss-of-function CRISPR screens in human lymphoma cell lines but are highly amenable for other experimental purposes.

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Acknowledgments

We thank all current and former colleagues who helped establish these protocols. This research was supported by the Intramural Research Program of the NIH, CCR, NCI. D.E.W. is a Damon Runyon Fellow (DRG-2208-14). S.R. is a H2020 Marie Sklodowska-Curie global fellow (#661066).

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Author notes

  1. Sandrine Roulland

    Present address: CNRS, INSERM, Centre d’Immunologie de Marseille-Luminy, Aix-Marseille Universite, Marseille, France

Authors and Affiliations

  1. Lymphoid Malignancies Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Daniel E. Webster, Sandrine Roulland & James D. Phelan

Authors
  1. Daniel E. Webster
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  2. Sandrine Roulland
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  3. James D. Phelan
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Correspondence to James D. Phelan .

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Editors and Affiliations

  1. Institute of Cell Biology (Cancer Research), Medical School, University of Duisburg-Essen, Nordrhein-Westfalen, Essen, Germany

    Ralf Küppers

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Webster, D.E., Roulland, S., Phelan, J.D. (2019). Protocols for CRISPR-Cas9 Screening in Lymphoma Cell Lines. In: Küppers, R. (eds) Lymphoma. Methods in Molecular Biology, vol 1956. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9151-8_16

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  • DOI: https://doi.org/10.1007/978-1-4939-9151-8_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9150-1

  • Online ISBN: 978-1-4939-9151-8

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