{
  "abstract": "CRISPR/Cas9 enables precision gene editing via homology-directed repair (HDR) for mutation correction and disease modelling. Here we present a customizable T cell single nucleotide variant (SNV) correction platform based on non-viral HDR, previously described in Mamia et al. Precision T Cell Correction Platform for Inborn Errors of Immunity. Molecular Therapy, 2025. To enable reproducible application, we provide a comprehensive, step-by-step 8-day HDR workflow for editing primary T cells, covering CRISPR/Cas9 reagent design, custom genomic editing from start to finish, optimization and validation strategies, on-target detection, and flow cytometric assessment of T cell phenotype and exhaustion. The platform separates locus-specific reagent optimization from a fixed cellular workflow, enabling implementation of new genomic targets without modifying the downstream T cell editing pipeline. Using this workflow, we have achieved up to 80% editing efficiency across multiple loci and donors. We have further optimized the T cell culture platform for PBMC stimulation to promote CD4+ and CD8+ T cell activation and proliferation. We validated this culture platform in 33 patients with Inborn Errors of Immunity using flow cytometry and observed that the defined culture conditions maintain low T cell exhaustion and support persistence of memory T cell populations. The protocol was developed under research-grade conditions but supports scalability and transition to preclinical and clinical GMP workflows. Overall, this adaptable platform is a broadly applicable framework for precision T cell engineering across diverse genomic targets offering opportunities for both in vitro disease modeling and therapeutic genome correction.",
  "abstract_shown": true,
  "affiliations": [
    "Norwegian Centre for Molecular Biosciences and Medicine, University of Oslo, Oslo, Norway",
    "Department of Pediatrics, Oslo University Hospital, Oslo, Norway",
    "Precision Immunotherapy Alliance, University of Oslo, Oslo, Norway",
    "School of Biosciences and Medicine, University of Surrey, Guildford, Surrey, United Kingdom",
    "Department of Pediatrics, Institute of Clinical Sciences, the Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden",
    "Department of Rheumatology and Inflammation Research, Institute of Medicine, the Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden",
    "Children’s Hospital and Pediatric Research Center, University of Helsinki and Helsinki University Hospital, Helsinki, Finland",
    "Clinical Microbiology, HUS Diagnostic Center, Helsinki University Hospital and University of Helsinki, Helsinki, Finland",
    "Translational Immunology Research Program, University of Helsinki, Helsinki, Finland",
    "Institute for Molecular Medicine Finland and Helsinki Institute of Life Science, University of Helsinki, Helsinki, Finland",
    "Endocrinology, Abdominal Center, Helsinki University Hospital, Helsinki, Finland",
    "University of Helsinki, ENDO-ERN (European Reference Network on Rare Endocrine Conditions), Helsinki, Finland",
    "Folkhälsan Institute of Genetics, and Stem Cells and Metabolism Research Program, University of Helsinki, Helsinki, Finland",
    "Institute of Clinical Medicine, University of Oslo, Oslo, Norway",
    "Section of Clinical Immunology and Infectious Diseases, Oslo University Hospital, Rikshospitalet, Oslo, Norway",
    "Pediatric Research Center, New Children’s Hospital, University of Helsinki and HUS Helsinki University Hospital, Helsinki, Finland",
    "Division of Hematology, Oncology, and Stem Cell Transplantation, New Children’s Hospital, HUS Helsinki University Hospital, Helsinki, Finland",
    "Inflammation Center, Department of Infectious Disease, Helsinki University Hospital and University of Helsinki, Helsinki, Finland"
  ],
  "article_type": "methods article",
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "affs": [
        0,
        1,
        2
      ],
      "name": "Katariina Mamia"
    },
    {
      "affs": [
        0,
        2,
        3
      ],
      "name": "Anniken Solveig Matheson Sollano"
    },
    {
      "affs": [
        0,
        4,
        5
      ],
      "name": "Vanja Lundberg"
    },
    {
      "affs": [
        6
      ],
      "name": "Svetlana Vakkilainen"
    },
    {
      "affs": [
        7,
        8,
        9
      ],
      "name": "Eliisa Kekäläinen"
    },
    {
      "affs": [
        10,
        11
      ],
      "name": "Camilla Schalin-Jäntti"
    },
    {
      "affs": [
        12
      ],
      "name": "Eira Leinonen"
    },
    {
      "affs": [
        13,
        14
      ],
      "name": "Børre Fevang"
    },
    {
      "affs": [
        8,
        15,
        16
      ],
      "name": "Juha Grönholm"
    },
    {
      "affs": [
        17
      ],
      "name": "Timi Martelius"
    },
    {
      "affs": [
        0,
        2
      ],
      "name": "Shiva Dahal-Koirala"
    },
    {
      "affs": [
        0,
        1,
        2
      ],
      "name": "Emma Maria Haapaniemi"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/82fx7d7pqzhqq.bib",
  "cited_by": 0,
  "doi": "10.3389/fgeed.2026.1817322",
  "fulltext_url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC13696951/",
  "id": "82fx7d7pqzhqq",
  "issns": [
    "2673-3439"
  ],
  "landing_url": "https://doi.org/10.3389/fgeed.2026.1817322",
  "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",
      "Methods"
    ],
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      "openalex",
      "1312|CRISPR and Genetic Engineering"
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  "pdf_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13696951/pdf/?tool=EBI",
  "pmcid": "PMC13696951",
  "pmid": "42936275",
  "published": "2026-09-30",
  "publisher": "Frontiers Media SA",
  "ris": "https://research.draconitic.com/paper/82fx7d7pqzhqq.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:13Z",
      "native_id": "PMC13696951",
      "source": "pmc",
      "url": "https://doi.org/10.3389/fgeed.2026.1817322"
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    {
      "first_seen": "2026-10-30T12:21:25Z",
      "native_id": "PMC:PMC13696951",
      "source": "europepmc",
      "url": "https://europepmc.org/article/PMC/PMC13696951"
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  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|CRISPR and Genetic Engineering",
      "subfield": "molecular-biology"
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  ],
  "title": "Protocol for non-viral HDR-based CRISPR/Cas9 platform for small custom editing in primary T cells",
  "title_text": "Protocol for non-viral HDR-based CRISPR/Cas9 platform for small custom editing in primary T cells",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:34:12Z",
  "url": "https://research.draconitic.com/paper/82fx7d7pqzhqq",
  "venue": "Frontiers in Genome Editing",
  "versions": []
}