Besides, the method of genetic perturbation of immune checkpoint genes has also been designed (Figure 3A); For example, recently a non-viral type of anti-CD19 CAR-T cell with PD1 integration through CRISPR-Cas9 has successfully developed, which showed superior ability to eradicate tumor cells in xenograft models and in adoptive therapy for eight R/R aggressive B cell NHL (NCT04213469) with a high rate (87.5%) of CR and durable responses without serious adverse events (113). The gene discussed is CD19; the disease is neoplasm.