TcBuster-M™ Enables Efficient Non-Viral Delivery of Multicistronic CD19-CAR Constructs in T and NK Cells 2259709

Abstract Introduction Viral vectors are widely used for immune cell engineering but pose challenges including limited cargo capacity and high production costs. TcBuster-M™, a transposase-based editing platform, offers a non-viral cell engineering alternative with broader cargo capacity and commercial availability. Methods Peripheral blood-derived T and NK cells were edited via electroporation with the TcBuster-M transposase and a multicistronic CD19-CAR transposon. Cells were expanded and assessed for cell growth and viability in addition to CAR expression and cell phenotype by flow cytometry. Genomic integration of the CD19-CAR was assessed by dPCR. Cytotoxicity was evaluated using a luciferase-based CD19+ target cell assay and cytokine secretion profile by Simple Plex (Ella). Results TcBuster-M-mediated cell engineering achieved high CD19-CAR expression in both T and NK cells while preserving cell viability and growth. Edited cells demonstrated potent, target-specific cytotoxicity and favorable cytokine secretion. Genomic analysis revealed stable integration with copy number variations below eight. Conclusion The TcBuster transposon system supports rapid, cost-effective cell manufacturing and enables delivery of complex therapeutic cargos, positioning it as a robust alternative to virus-mediated editing systems for immunotherapy development. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)