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Inducing multiple nicks promotes interhomolog homologous recombination to correct heterozygous mutations in somatic cells

Author

Listed:
  • Akiko Tomita

    (Osaka University, Suita)

  • Hiroyuki Sasanuma

    (Tokyo Metropolitan Institute of Medical Science)

  • Tomoo Owa

    (Osaka University, Suita)

  • Yuka Nakazawa

    (Nagoya University
    Nagoya University Graduate School of Medicine)

  • Mayuko Shimada

    (Nagoya University
    Nagoya University Graduate School of Medicine)

  • Takahiro Fukuoka

    (Nagoya University
    Genomedia Inc.)

  • Tomoo Ogi

    (Nagoya University
    Nagoya University Graduate School of Medicine)

  • Shinichiro Nakada

    (Osaka University, Suita
    Osaka University, Suita)

Abstract

CRISPR/Cas9-mediated gene editing has great potential utility for treating genetic diseases. However, its therapeutic applications are limited by unintended genomic alterations arising from DNA double-strand breaks and random integration of exogenous DNA. In this study, we propose NICER, a method for correcting heterozygous mutations that employs multiple nicks (MNs) induced by Cas9 nickase and a homologous chromosome as an endogenous repair template. Although a single nick near the mutation site rarely leads to successful gene correction, additional nicks on homologous chromosomes strongly enhance gene correction efficiency via interhomolog homologous recombination (IH-HR). This process partially depends on BRCA1 and BRCA2, suggesting the existence of several distinct pathways for MN-induced IH-HR. According to a genomic analysis, NICER rarely induces unintended genomic alterations. Furthermore, NICER restores the expression of disease-causing genes in cells derived from genetic diseases with compound heterozygous mutations. Overall, NICER provides a precise strategy for gene correction.

Suggested Citation

  • Akiko Tomita & Hiroyuki Sasanuma & Tomoo Owa & Yuka Nakazawa & Mayuko Shimada & Takahiro Fukuoka & Tomoo Ogi & Shinichiro Nakada, 2023. "Inducing multiple nicks promotes interhomolog homologous recombination to correct heterozygous mutations in somatic cells," Nature Communications, Nature, vol. 14(1), pages 1-20, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-41048-5
    DOI: 10.1038/s41467-023-41048-5
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