Generation of Murine Chimeric Antigen Receptor T Cells for Adoptive T Cell Therapy for Melanoma

Expression of chimeric antigen receptors enables T cells to be redirected toward melanoma-associated antigens in an MHC-independent manner. Retroviral transduction can be used to introduce these receptor constructs into murine CD4⁺ and CD8⁺ T cells. Here, we outline the generation of retroviral supernatants and the activation, transduction, expansion, and selection of murine T cells expressing the chimeric-PD1 receptor. This protocol can be adapted for any murine chimeric antigen receptor construct.