{
  "abstract": "Chimeric antigen receptor (CAR) T cell therapy has revolutionized the treatment of B cell malignancies, and extending this success to other cancers remains an urgent clinical objective. CRISPR activation (CRISPRa) is a gene nomination platform with the potential to direct next-generation cell product development, but genome-wide surveys using this technology in primary T cells have been limited by prohibitive cell requirements and short-lived activity. Here, we describe a CAR-restricted T cell expansion platform generating yields of CRISPRa-CAR T cells far exceeding those required for genome-scale screening (∼10⁹). CRISPRa activity is sustained by incorporating divergent, duplicate elongation factor 1α core/human T cell leukemia virus hybrid promoters and by delivering guide RNAs late by lentiviral transduction to avoid losing perturbations that compromise early T cell activation or expansion. Manufactured CRISPRa-CAR T cells maintain potent on-target gene overexpression (>85% target⁺) across varied cell subsets, including those associated with T cell exhaustion, and over extended culture periods. When applied to survival-based genome-wide screens, CRISPRa-CAR T cell pools nominate endogenous genes that enhance CAR T cell persistence across varied cytokine levels in the presence of tumor (e.g., IRF4, GFI1, and GSE1). In a proof-of-concept study, we find that the survival effect of GSE1 can be preserved using a clinically relevant transgene overexpression strategy. More broadly, this screening framework establishes a foundation for systematic identification and downstream validation of genes that enhance cellular therapies.",
  "abstract_shown": true,
  "affiliations": [
    "Seattle Children's Therapeutics, Seattle Children's Research Institute, 1920 Terry Ave, Seattle, WA 98101, USA.",
    "Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA.",
    "Ben Towne Center for Childhood Cancer and Blood Disorders Research, Seattle Children's Research Institute, 1920 Terry Ave, Seattle, WA 98101, USA.",
    "Department of Bioengineering, University of Washington, 3720 15th Ave NE, Seattle, WA 98195, USA.",
    "Division of Hematology, Oncology, and Bone Marrow Transplant, Department of Pediatrics, University of Washington Medical School, 4800 Sand Point Way NE, Seattle, WA 98105, USA."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 19,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Benjamin C Curtis"
    },
    {
      "affs": [
        0
      ],
      "name": "Cailyn H Spurrell"
    },
    {
      "affs": [
        0
      ],
      "name": "Lindsay Flint"
    },
    {
      "affs": [
        0
      ],
      "name": "Aalton M Lande",
      "orcid": "0009-0000-8613-4939"
    },
    {
      "affs": [
        0
      ],
      "name": "Marissa Leonardi"
    },
    {
      "affs": [
        0
      ],
      "name": "James M Rosser",
      "orcid": "0009-0009-4744-1165"
    },
    {
      "affs": [
        0
      ],
      "name": "Tavie Parker"
    },
    {
      "affs": [
        0
      ],
      "name": "Tiffanie Chai"
    },
    {
      "affs": [
        0
      ],
      "name": "Ardizon Valdez"
    },
    {
      "affs": [
        2
      ],
      "name": "Ryma Toumi"
    },
    {
      "affs": [
        0
      ],
      "name": "Nat Murren"
    },
    {
      "affs": [
        0
      ],
      "name": "Michael Fitzgerald"
    },
    {
      "affs": [
        0
      ],
      "name": "Jasmin Martinez-Reyes"
    },
    {
      "affs": [
        0
      ],
      "name": "Finn Watt"
    },
    {
      "affs": [
        0,
        3
      ],
      "name": "Christopher P Saxby"
    },
    {
      "affs": [
        0
      ],
      "name": "Corinne Summers"
    },
    {
      "affs": [
        0
      ],
      "name": "Kamila S Gwiazda"
    },
    {
      "affs": [
        0
      ],
      "name": "Michael C V Jensen"
    },
    {
      "affs": [
        2,
        4
      ],
      "name": "Shannon K Oda"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/0cf0ps16tfnen.bib",
  "cited_by": 0,
  "doi": "10.1016/j.omtn.2026.103117",
  "fulltext_url": "https://europepmc.org/articles/PMC13694873",
  "id": "0cf0ps16tfnen",
  "issns": [
    "2162-2531"
  ],
  "landing_url": "https://europepmc.org/article/MED/42927201",
  "language": "eng",
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "2730|CAR-T cell therapy research"
    ]
  ],
  "openalex": "W7214706168",
  "pdf_url": "https://europepmc.org/articles/PMC13694873?pdf=render",
  "pmcid": "PMC13694873",
  "pmid": "42927201",
  "published": "2026-09-28",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/0cf0ps16tfnen.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:21:18Z",
      "native_id": "MED:42927201",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42927201"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "openalex:2730|CAR-T cell therapy research",
      "subfield": "oncology"
    }
  ],
  "title": "Genome-wide CRISPRa screens identify modulators of CAR T survival across diverse tumor cytokine milieus.",
  "title_text": "Genome-wide CRISPRa screens identify modulators of CAR T survival across diverse tumor cytokine milieus.",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:21:18Z",
  "url": "https://research.draconitic.com/paper/0cf0ps16tfnen",
  "venue": "Molecular therapy. Nucleic acids",
  "versions": []
}