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Pooled library screening with multiplexed Cpf1 library

Author

Listed:
  • Jintan Liu

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

  • Sanjana Srinivasan

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

  • Chieh-Yuan Li

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

  • I-Lin Ho

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

  • Johnathon Rose

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

  • MennatAllah Shaheen

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

  • Gang Wang

    (The University of Texas MD Anderson Cancer Center)

  • Wantong Yao

    (The University of Texas MD Anderson Cancer Center
    The University of Texas MD Anderson Cancer Center)

  • Angela Deem

    (The University of Texas MD Anderson Cancer Center)

  • Chris Bristow

    (The University of Texas MD Anderson Cancer Center)

  • Traver Hart

    (The University of Texas MD Anderson Cancer Center
    The University of Texas MD Anderson Cancer Center)

  • Giulio Draetta

    (The University of Texas MD Anderson Cancer Center
    MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Houston)

Abstract

Capitalizing on the inherent multiplexing capability of AsCpf1, we developed a multiplexed, high-throughput screening strategy that minimizes library size without sacrificing gene targeting efficiency. We demonstrated that AsCpf1 can be used for functional genomics screenings and that an AsCpf1-based multiplexed library performs similarly as compared to currently available monocistronic CRISPR/Cas9 libraries, with only one vector required for each gene. We construct the smallest whole-genome CRISPR knock-out library, Mini-human, for the human genome (n = 17,032 constructs targeting 16,977 protein-coding genes), which performs favorably compared to conventional Cas9 libraries.

Suggested Citation

  • Jintan Liu & Sanjana Srinivasan & Chieh-Yuan Li & I-Lin Ho & Johnathon Rose & MennatAllah Shaheen & Gang Wang & Wantong Yao & Angela Deem & Chris Bristow & Traver Hart & Giulio Draetta, 2019. "Pooled library screening with multiplexed Cpf1 library," Nature Communications, Nature, vol. 10(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-10963-x
    DOI: 10.1038/s41467-019-10963-x
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