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High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening

Author

Listed:
  • Rodrigo A. Gier

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

  • Krista A. Budinich

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

  • Niklaus H. Evitt

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

  • Zhendong Cao

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

  • Elizabeth S. Freilich

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

  • Qingzhou Chen

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

  • Jun Qi

    (Harvard Medical School)

  • Yemin Lan

    (University of Pennsylvania)

  • Rahul M. Kohli

    (University of Pennsylvania
    University of Pennsylvania)

  • Junwei Shi

    (University of Pennsylvania
    University of Pennsylvania
    University of Pennsylvania)

Abstract

CRISPR-based genetic screening has revolutionized cancer drug target discovery, yet reliable, multiplex gene editing to reveal synergies between gene targets remains a major challenge. Here, we present a simple and robust CRISPR-Cas12a-based approach for combinatorial genetic screening in cancer cells. By engineering the CRISPR-AsCas12a system with key modifications to the Cas protein and its CRISPR RNA (crRNA), we can achieve high efficiency combinatorial genetic screening. We demonstrate the performance of our optimized AsCas12a (opAsCas12a) through double knockout screening against epigenetic regulators. This screen reveals synthetic sick interactions between Brd9&Jmjd6, Kat6a&Jmjd6, and Brpf1&Jmjd6 in leukemia cells.

Suggested Citation

  • Rodrigo A. Gier & Krista A. Budinich & Niklaus H. Evitt & Zhendong Cao & Elizabeth S. Freilich & Qingzhou Chen & Jun Qi & Yemin Lan & Rahul M. Kohli & Junwei Shi, 2020. "High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening," Nature Communications, Nature, vol. 11(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-17209-1
    DOI: 10.1038/s41467-020-17209-1
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    Cited by:

    1. Natasha Ramakrishnan & Tyler M. Weaver & Lindsey N. Aubuchon & Ayda Woldegerima & Taylor Just & Kevin Song & Alessandro Vindigni & Bret D. Freudenthal & Priyanka Verma, 2024. "Nucleolytic processing of abasic sites underlies PARP inhibitor hypersensitivity in ALC1-deficient BRCA mutant cancer cells," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    2. Ruitong Li & Olaf Klingbeil & Davide Monducci & Michael J. Young & Diego J. Rodriguez & Zaid Bayyat & Joshua M. Dempster & Devishi Kesar & Xiaoping Yang & Mahdi Zamanighomi & Christopher R. Vakoc & Ta, 2022. "Comparative optimization of combinatorial CRISPR screens," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
    3. Guanhua Xun & Zhixin Zhu & Nilmani Singh & Jingxia Lu & Piyush K. Jain & Huimin Zhao, 2024. "Harnessing noncanonical crRNA for highly efficient genome editing," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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