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- W2563619005 abstract "Genetically engineered cancer targeting CAR-T cells have generated promising results in a series of B cell derived cancers. Development of these therapies for solid tumor applications has been much more difficult and will likely require extensive T cell engineering to improve both efficacy and safety. Advanced gene editing approaches were developed to enable simultaneous disruption of a target gene combined with introduction of exogenous transgenes at the disrupted locus. A gene specific megaTAL nuclease was used to generate double stranded DNA breaks followed by transduction with adeno-associated virus (AAV) encoding new genetic information flanked by regions of homology proximal to the nuclease breakpoint. Highly efficient introduction of a CD19-specific CAR transgene into the T cell receptor-alpha constant (TRAC) locus was demonstrated using this approach. T cells treated with the TRAC megaTAL and corresponding AAV encoding a CD19 CAR and TRAC homology arms generated greater than 50% of CD19-CAR positive T cells that no longer expressed the T cell receptor complex. In vitro assays confirmed that TRAC-targeted CD19-CAR T cells were comparable to CD19 CAR-T cells generated by lentiviral transduction in their cytotoxicity and cytokine responses against CD19+ Nalm-6 cells. These findings demonstrate the potential of megaTAL driven homology directed T cell genome engineering to obviate the need for traditional integrating viral vectors and generate a defined and potentially more potent T cell product by combining gene disruption with targeted transgene integration." @default.
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- W2563619005 date "2016-05-01" @default.
- W2563619005 modified "2023-10-14" @default.
- W2563619005 title "323. Efficient Generation of CART Cells by Homology Directed Transgene Integration into the TCR-Alpha Locus" @default.
- W2563619005 doi "https://doi.org/10.1016/s1525-0016(16)33132-x" @default.
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