Evidence map›Paper›PMID 39815622›Full record

ArticleMolecular therapy : the journal of the American Society of Gene Therapy2025

Virus-free CRISPR knockin of a chimeric antigen receptor into KLRC1 generates potent GD2-specific natural killer cells.

Keerthana Shankar, Isabelle Zingler-Hoslet, Diana M Tabima, Seth Zima, Lei Shi, Kirstan Gimse, Matthew H Forsberg, Varun Katta, Sage Z Davis, Daniel Maldonado and 6 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Molecular therapy : the journal of the American Society of Gene Therapy, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.

0numbers the graph read from it
0cells of the map it votes in
12citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

12 citing papers in PubMed.

  1. Article
  2. Article
  3. Review
  4. The CD94/NKG2A-HLA-E Axis as a Target in Cancer Immunotherapy: A Critical Perspective.Clinical cancer research : an official journal of the American Association for Cancer Research · 2026
    Review
  5. Review
  6. Review
  7. Review
  8. Article
  9. Review
  10. Rendering NK Cells Antigen-Specific for the Therapy of Solid Tumours.International journal of molecular sciences · 2025
    Review
  11. Review
  12. Review
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

16 authors.

Keerthana ShankarDepartment of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
Isabelle Zingler-HosletDepartment of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
Diana M TabimaWisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
Seth ZimaDepartment of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Dermatology, University of Wisconsin-Madison, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI 53715, USA.
Lei ShiDepartment of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI 53715, USA.
Kirstan GimseWisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
Matthew H ForsbergDepartment of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI 53715, USA.
Varun KattaDepartment of Hematology, St Jude Children's Research Hospital, Memphis, TN 38105, USA.
Sage Z DavisCenter for Human Genomics and Precision Medicine, University of Wisconsin-Madison, Madison, WI 53705, USA.
Daniel MaldonadoWisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, WI 53706, USA.
Brittany E RussellDepartment of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
Muhammed MurtazaCenter for Human Genomics and Precision Medicine, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Surgery, University of Wisconsin-Madison, Madison, WI 53792, USA.
Shengdar Q TsaiDepartment of Hematology, St Jude Children's Research Hospital, Memphis, TN 38105, USA.
Jose M AyusoDepartment of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Dermatology, University of Wisconsin-Madison, Madison, WI 53715, USA; Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI 53715, USA.
Christian M CapitiniDepartment of Dermatology, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53715, USA.
Krishanu SahaDepartment of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53715, USA. Electronic address: ksaha@wisc.edu.

Funding

UW COMPREHENSIVE CANCER CENTER SUPPORTP30CA014520 · NCI · UNIVERSITY OF WISCONSIN-MADISON · PI Justine Yang Bruce · 1985 to 2026
$142.6M
Assembly of Novel Gene Editing Particles to Understand Genome Surgery in Patient-Derived CellsR35GM119644 · NIGMS · UNIVERSITY OF WISCONSIN-MADISON · PI SAHA, KRISHANU · 2016 to 2025
$4.0M
Label-free imaging of CAR T cell metabolismR01CA278051 · NCI · MORGRIDGE INSTITUTE FOR RESEARCH, INC. · PI Christian Capitini, Krishanu Saha · 2023 to 2026
$2.6M
Ultra-sensitive, unbiased, high-throughput, biochemical CHANGE-seq genome-wide activity and gRNA sequencing assays for therapeutic genome editing INDsU01AI176471 · NIAID · ST. JUDE CHILDREN'S RESEARCH HOSPITAL · PI TSAI, SHENGDAR · 2023 to 2025
$1.4M
Multi-color Benchtop Flow CytometerS10RR025483 · NCRR · UNIVERSITY OF WISCONSIN-MADISON · PI BUSHMAN, WADE A · 2009 to 2009
$449k
NCI NIH HHS P30 CA014520NCI NIH HHS R01 CA278051NCRR NIH HHS S10 RR025483NIAID NIH HHS U01 AI176471NIGMS NIH HHS R35 GM119644
6 · The paper itself

Abstract

Natural killer (NK) cells are an appealing off-the-shelf, allogeneic cellular therapy due to their cytotoxic profile. However, their activity against solid tumors remains suboptimal in part due to the upregulation of NK-inhibitory ligands, such as HLA-E, within the tumor microenvironment. Here, we utilize CRISPR-Cas9 to disrupt the KLRC1 gene (encoding the HLA-E-binding NKG2A receptor) and perform non-viral insertion of a GD2-targeting chimeric antigen receptor (CAR) within NK cells isolated from human peripheral blood. Genome editing with CRISPR-Cas9 ribonucleoprotein complexes yields efficient genomic disruption of the KLRC1 gene with 98% knockout efficiency and specific knockin of the GD2 CAR transgene as high as 23%, with minimal off-target activity as shown by CHANGE-seq, in-out PCR, amplicon sequencing, and long-read whole-genome sequencing. KLRC1-GD2 CAR NK cells display high viability and proliferation, as well as precise cellular targeting and potency against GD2

Indexed as

CRISPR-Cas SystemsKiller Cells, NaturalReceptors, Chimeric AntigenCell Line, TumorGangliosidesGene EditingGene Knock-In TechniquesHumansImmunotherapy, Adoptiveganglioside, GD2GangliosidesReceptors, Chimeric AntigenCARCRISPR-Cas9GD2KLRC1NK cellsNKG2Anon-viral

Identifiers

PMID39815622
PMCPMC11897758

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.