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Next-Generation Sequencing of Genome-Wide CRISPR Screens

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Next Generation Sequencing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1712))

Abstract

Genome-wide functional genomic screens utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system have proven to be a powerful tool for systematic genomic perturbation in mammalian cells and provide an alternative to previous screens utilizing RNA interference technology. The wide availability of these libraries through public plasmid repositories as well as the decreasing cost and speed in quantifying these screens using high-throughput next-generation sequencing (NGS) allows for the adoption of the technology in a variety of laboratories interested in diverse biologic questions. Here, we describe the protocol to generate next-generation sequencing libraries from genome-wide CRISPR genomic screens.

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Acknowledgments

We thank the Rana lab members, John Shimishata, and Steven Head at The Scripps Research Institute Genomic Core. This work was supported in part by grants from the National Institutes of Health to T.M.R., E.Y, is supported by the National Cancer Institute of the National Institutes of Health under award number T32CA121938. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Author information

Authors and Affiliations

  1. Division of Hematology-Oncology, Department of Internal Medicine, University of California, San Diego, La Jolla, CA, 92037, USA

    Edwin H. Yau

  2. Solid Tumor Therapeutics Program, Moores Cancer Center, University of California, San Diego, 3855 Health Sciences Drive, La Jolla, CA, 92093, USA

    Edwin H. Yau & Tariq M. Rana

  3. Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0762, La Jolla, CA, 92093, USA

    Tariq M. Rana

  4. Institute for Genomic Medicine, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, 92093, USA

    Tariq M. Rana

Authors
  1. Edwin H. Yau
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  2. Tariq M. Rana
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Corresponding author

Correspondence to Tariq M. Rana .

Editor information

Editors and Affiliations

  1. Next Generation Sequencing and Microarray Core Facility, The Scripps Research Institute, La Jolla, California, USA

    Steven R. Head

  2. Next Generation Sequencing and Microarray Core Facility, The Scripps Research Institute, La Jolla, California, USA

    Phillip Ordoukhanian

  3. Department of Molecular Medicine, The Scripps Research Institute Center for Organ and Cell Transplantation, La Jolla, California, USA

    Daniel R. Salomon

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Yau, E.H., Rana, T.M. (2018). Next-Generation Sequencing of Genome-Wide CRISPR Screens. In: Head, S., Ordoukhanian, P., Salomon, D. (eds) Next Generation Sequencing. Methods in Molecular Biology, vol 1712. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7514-3_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7514-3_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7512-9

  • Online ISBN: 978-1-4939-7514-3

  • eBook Packages: Springer Protocols

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