Generation of Chimeric Antigen Receptor T Cells Using Gammaretroviral Vectors
Manufacturing chimeric antigen receptor (CAR)-modified T cells requires incorporation of the CAR transgene, for which viral vectors are most often used. Here, we describe the generation of CAR T cells using primary human T cells and a non–self-inactivating gammaretroviral vector encoding a CAR transgene. The gammaretroviral vector is produced by 293T cells transiently transfected with DNA plasmids encoding necessary components of the viral vector. The resulting viral particles efficiently infect activated T cells and integrate the CAR transgene into the genome of dividing cells for stable expression.
Key wordsGammaretroviral vectors Vector production CAR T cell generation T cell transduction Chimeric antigen receptors
The authors thank Catherine Gillespie for editing the chapter. This work was supported by the CPRIT Awards RP180810 and RP150611, and by the Leukemia and Lymphoma Society TRP Award.
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