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Characterizing and controlling CRISPR repair outcomes in nondividing human cells

Author

Listed:
  • Gokul N. Ramadoss

    (Gladstone Institutes
    University of California)

  • Samali J. Namaganda

    (Gladstone Institutes)

  • Manasi M. Kumar

    (Gladstone Institutes)

  • Jennifer R. Hamilton

    (University of California
    University of California)

  • Rohit Sharma

    (University of California
    University of California)

  • Karena G. Chow

    (Gladstone Institutes)

  • Luke A. Workley

    (Gladstone Institutes
    University of California)

  • Bria L. Macklin

    (Gladstone Institutes)

  • Mengyuan Sun

    (Gladstone Institutes)

  • Alvin S. Ha

    (Gladstone Institutes
    University of California)

  • Jia-Cheng Liu

    (NIH)

  • Christof Fellmann

    (Gladstone Institutes
    University of California)

  • Hannah L. Watry

    (Gladstone Institutes)

  • Philip H. Dierks

    (Gladstone Institutes)

  • Rudra S. Bose

    (University of California)

  • Julianne Jin

    (University of California)

  • Barbara S. Perez

    (University of California
    University of California)

  • Cindy R. Sandoval Espinoza

    (University of California
    University of California)

  • Madeline P. Matia

    (Gladstone Institutes)

  • Serena H. Lu

    (Gladstone Institutes)

  • Luke M. Judge

    (Gladstone Institutes
    University of California)

  • Brian R. Shy

    (Gladstone Institutes
    University of California
    University of California)

  • Andre Nussenzweig

    (NIH)

  • Britt Adamson

    (Princeton University
    Princeton University)

  • Niren Murthy

    (University of California
    University of California)

  • Jennifer A. Doudna

    (Gladstone Institutes
    University of California
    University of California
    University of California)

  • Martin Kampmann

    (University of California
    University of California)

  • Bruce R. Conklin

    (Gladstone Institutes
    University of California
    University of California
    University of California)

Abstract

Genome editing is poised to revolutionize treatment of genetic diseases, but poor understanding and control of DNA repair outcomes hinders its therapeutic potential. DNA repair is especially understudied in nondividing cells like neurons, limiting the efficiency and precision of genome editing in many clinically relevant tissues. Here, we address this barrier by using induced pluripotent stem cells (iPSCs) and iPSC-derived neurons to examine how postmitotic human neurons repair Cas9-induced DNA damage. CRISPR editing outcomes differ dramatically in neurons compared to genetically identical dividing cells: neurons take longer to fully resolve this damage, and upregulate non-canonical DNA repair factors in the process. Manipulating this response with chemical or genetic perturbations allows us to direct DNA repair toward desired editing outcomes in nondividing human neurons, cardiomyocytes, and primary T cells. By studying DNA repair in clinically relevant cells, we reveal unforeseen challenges and opportunities for precise therapeutic editing.

Suggested Citation

  • Gokul N. Ramadoss & Samali J. Namaganda & Manasi M. Kumar & Jennifer R. Hamilton & Rohit Sharma & Karena G. Chow & Luke A. Workley & Bria L. Macklin & Mengyuan Sun & Alvin S. Ha & Jia-Cheng Liu & Chri, 2025. "Characterizing and controlling CRISPR repair outcomes in nondividing human cells," Nature Communications, Nature, vol. 16(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-66058-3
    DOI: 10.1038/s41467-025-66058-3
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    References listed on IDEAS

    as
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    Full references (including those not matched with items on IDEAS)

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