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shRNA-mediated TCR knockdown as a foundation for allogeneic CAR19 T-cells generated by single-step genetic modification with the piggyBac transposase

  • Author Footnotes
    * Corresponding author.
    D. Bishop
    Footnotes
    * Corresponding author.
    Affiliations
    Westmead Institute for Medical Research, Westmead, New South Wales, Australia

    Haematology Department, Westmead Hospital, Westmead, New South Wales, Australia

    Sydney Medical School, The University of Sydney, Sydney, New South Wales, Australia
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  • D. Gottlieb
    Affiliations
    Westmead Institute for Medical Research, Westmead, New South Wales, Australia

    Haematology Department, Westmead Hospital, Westmead, New South Wales, Australia

    Sydney Medical School, The University of Sydney, Sydney, New South Wales, Australia
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  • K. Micklethwaite
    Affiliations
    Westmead Institute for Medical Research, Westmead, New South Wales, Australia

    Haematology Department, Westmead Hospital, Westmead, New South Wales, Australia

    Sydney Medical School, The University of Sydney, Sydney, New South Wales, Australia
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  • Author Footnotes
    * Corresponding author.
      Aim: Autologous CD19-specific chimeric antigen receptor (CAR19) T cells have shown remarkable efficacy against relapsed/refractory B cell malignancies. However, manufacture of individualised products using viral vectors is expensive, delays treatment, is unfeasible in patients with insufficient healthy T cells, and risks contamination with CD19neg malignant B cells that may inadvertently be genetically modified to express CAR. We sought to evaluate short hairpin RNA (shRNA) suppression of the T cell receptor (TCR) in low-cost piggyBac-generated CAR T cells, in order to generate a ready-made allogeneic product devoid of alloreactivity.
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