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oai:figshare.com:article/34054605 2026-10-02T05:37:32Z category_696 portal_316 item_type_3 month_year_10_2026

Data Sheet 1_Split intein-mediated reconstitution of regulatable CAR T-cells.pdf Vladimir Mulens-Arias (8702370) Marco Archinti (25162668) Miriam Bastida (25162671) Nuria Martínez (25162674) Eleonora Pinto (3418427) Víctor M. Díaz (9184031) Pablo Menéndez (5662426) Genetic Immunology CAR T-cells immunotherapy modular protein trans-splicing split inteins Introduction<p>Chimeric antigen receptor (CAR) T-cell therapy has transformed the treatment of B-cell malignancies. However, strategies that improve the modularity and controllability of CAR signaling could broaden its applicability and safety across additional clinical contexts. Here, we describe a regulable CAR T-cell platform based on split intein-mediated protein trans-splicing (PTS) to conditionally reconstitute functional receptors in primary T-cells.</p>Methods<p>As a proof of concept, we engineered a split CAR targeting CD1a (NCT05679895), a surface antigen expressed in T-cell acute lymphoblastic leukemia (T-ALL). The system consists of a membrane-anchored C-terminal scaffold containing the IMPDH.1^C split intein fused to 4-1BB and CD3ζ signaling domains, and a soluble N-terminal adaptor composed of an anti-CD1a single-chain variable fragment (scFv) fused to the complementary IMPDH.1^N split intein. </p>Results<p>Upon delivery of the adaptor module, the intein fragments undergo PTS, generating a functional membrane-bound CD1a CAR with splicing efficiencies of 85%. Reconstituted CAR T-cells exhibited antigen-specific cytotoxicity against CD1a⁺ T-ALL cells coupled to high levels of interferon-γ secretion. </p>Discussion<p>These findings demonstrate that split intein-mediated CAR reconstitution enables a modular platform for conditional CAR reconstitution and transient control of receptor availability. </p> 2026-10-02T05:37:32Z Dataset Dataset 10.3389/fimmu.2026.1844972.s001 https://figshare.com/articles/dataset/Data_Sheet_1_Split_intein-mediated_reconstitution_of_regulatable_CAR_T-cells_pdf/34054605 CC BY 4.0