In this study, to address these challenges, we utilized RN-coated plastic bag preloading with low-temperature shaking (LTS) and performed large-scale T cell manufacturing of both endoribonuclease MazF-modified CD4+ T cells for HIV-1 gene therapy and MAGE-A4 TCR-modified T cells for MAGE-A4-antigen-expressing cancer gene therapy. The gene discussed is CD4; the disease is cancer.