{
  "abstract": "Precise genome editing in hematopoietic stem cells (HSCs) holds great potential for treating inherited blood disorders; however, low knock-in (KI) efficiency and loss of functional HSCs after genome editing remain major obstacles. Here, we demonstrate that transient inhibition of Ataxia-Telangiectasia Mutated (ATM) kinase markedly enhanced KI efficiency in mouse HSCs genome-edited with Cas9/ribonucleoprotein (RNP) and AAV donor DNA. Phosphoproteomic and capillary western blot analyses revealed that ATM inhibition suppressed aberrant Cas9-AAV-induced ATM activation and the subsequent DNA damage response, reduced p53-dependent apoptosis, and enhanced survival of KI HSCs. In serial transplantation assays, ATM inhibition preserved long-term engrafting KI HSCs, increasing their frequency from ~0.3 to 40% in secondary recipients, an over 100-fold improvement. Furthermore, in an X-linked severe combined immunodeficient mouse model, ATM inhibition increased KI efficiency and restored T- and B cell differentiation. Comprehensive safety analyses, including long-read nanopore sequencing, amplicon deep sequencing, whole genome sequencing and clonal tracking, did not detect increased genomic alterations, chromosomal abnormalities, or clonal expansion following transient ATM inhibition. Taken together, these findings establish transient ATM inhibition as an effective strategy to enhance KI-mediated genome editing while maintaining the long-term repopulating capacity of HSCs.",
  "abstract_shown": true,
  "affiliations": [
    "Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, Shimotsuke 329-0498, Tochigi, Japan",
    "Laboratory of Regenerative and Cellular Medicine, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsuke 329-0498, Tochigi, Japan",
    "Division of Animal Resource Development, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsuke 329-0498, Tochigi, Japan",
    "Center for Experimental Medicine, Jichi Medical University, Shimotsuke 329-0498, Tochigi, Japan",
    "Department of Biochemistry, School of Medicine, Jichi Medical University, Shimotsuke 329-0498, Tochigi, Japan",
    "San Raffaele Telethon Institute for Gene Therapy, Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele Scientific Institute, Milan 20132, Italy",
    "Faculty of Medicine and Surgery, Faculty of Medicine and Surgery, Vita-Salute San Raffaele University, Milan 20132, Italy",
    "Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, IL 60612",
    "Translational Research Laboratory, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsuke 329-0498, Tochigi, Japan"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Munkh-Erdene Natsagdorj",
      "orcid": "0009-0004-9658-8167"
    },
    {
      "affs": [
        0,
        1,
        2,
        3
      ],
      "name": "Hiromasa Hara",
      "orcid": "0000-0003-1433-1241"
    },
    {
      "affs": [
        0
      ],
      "name": "Kohei Iida"
    },
    {
      "affs": [
        4
      ],
      "name": "Yuji Kashiwakura"
    },
    {
      "affs": [
        4
      ],
      "name": "Nemekhbayar Baatartsogt"
    },
    {
      "affs": [
        0
      ],
      "name": "Enkhzaya Ganzorig",
      "orcid": "0009-0002-6184-3127"
    },
    {
      "affs": [
        5,
        6
      ],
      "name": "Daniele Canarutto",
      "orcid": "0000-0001-6999-8479"
    },
    {
      "affs": [
        7
      ],
      "name": "Sandra Pinho",
      "orcid": "0000-0002-5241-7364"
    },
    {
      "affs": [
        5,
        6
      ],
      "name": "Luigi Naldini"
    },
    {
      "affs": [
        4
      ],
      "name": "Tsukasa Ohmori"
    },
    {
      "affs": [
        0,
        1,
        8
      ],
      "name": "Yutaka Hanazono",
      "orcid": "0000-0003-4610-7770"
    },
    {
      "affs": [
        0
      ],
      "name": "Fumio Nakahara",
      "orcid": "0000-0003-1515-8284"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/1n0y9h3034dwp.bib",
  "cited_by": 0,
  "doi": "10.1073/pnas.2531364123",
  "fulltext_url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC13715756/",
  "id": "1n0y9h3034dwp",
  "issns": [
    "0027-8424",
    "1091-6490"
  ],
  "landing_url": "https://doi.org/10.1073/pnas.2531364123",
  "language": "en",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "403"
    ],
    [
      "heading",
      "Applied Biological Sciences"
    ],
    [
      "heading",
      "Biological Sciences"
    ],
    [
      "openalex",
      "1312|CRISPR and Genetic Engineering"
    ]
  ],
  "openalex": "W7223040400",
  "pdf_url": null,
  "pmcid": "PMC13715756",
  "pmid": "42900716",
  "published": "2026-10-20",
  "publisher": "National Academy of Sciences",
  "ris": "https://research.draconitic.com/paper/1n0y9h3034dwp.ris",
  "sources": [
    {
      "first_seen": "2026-11-01T12:22:58Z",
      "native_id": "PMC13715756",
      "source": "pmc",
      "url": "https://doi.org/10.1073/pnas.2531364123"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|CRISPR and Genetic Engineering",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Transient ATM inhibition enhances knock-in efficiency in hematopoietic stem cells by attenuating the DNA damage response",
  "title_text": "Transient ATM inhibition enhances knock-in efficiency in hematopoietic stem cells by attenuating the DNA damage response",
  "type": "peer-reviewed",
  "updated_at": "2026-11-01T12:22:58Z",
  "url": "https://research.draconitic.com/paper/1n0y9h3034dwp",
  "venue": "Proceedings of the National Academy of Sciences of the United States of America",
  "versions": []
}