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Optimized gene engineering of murine CAR-T cells reveals the beneficial effects of IL-15 coexpression

J Exp Med. 2021-02; 
Evripidis Lanitis, Giorgia Rota, Paris Kosti, Catherine Ronet, Aodrenn Spill, Bili Seijo, Pedro Romero, Denarda Dangaj, George Coukos, Melita Irving
Products/Services Used Details Operation
PCR and Cloning … Primers to specifically amplify regions of the DC101 scFv of the CAR cassette, or the mIL-15 transgene, were designed using the GenScript … PCR amplification of the housekeeping gene GAPDH was performed as a control, and to allow normalization of samples. The following … Get A Quote

摘要

Limited clinical benefit has been demonstrated for chimeric antigen receptor (CAR) therapy of solid tumors, but coengineering strategies to generate so-called fourth-generation (4G) CAR-T cells are advancing toward overcoming barriers in the tumor microenvironment (TME) for improved responses. In large part due to technical challenges, there are relatively few preclinical CAR therapy studies in immunocompetent, syngeneic tumor-bearing mice. Here, we describe optimized methods for the efficient retroviral transduction and expansion of murine T lymphocytes of a predominantly central memory T cell (TCM cell) phenotype. We present a bicistronic retroviral vector encoding both a tumor vasculature-targeted CAR and mu... More

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