To produce piggyBac-CD19 CAR-T cells, we used 4D-Nucleofector device and P3 Primary Cell 4D-Nucleofector X Kit (Lonza, Basel,
Switzerland) for gene transfer. Briefly, we used program EO-115 to electroporate 1 × 107 PBMC with 5 µg each of pIRII-CAR.CD19 transposon plasmid and pCMV-piggyBac transposase plasmid. Transfection efficiency figure 2a, 30-55%, comparison with retriviruses and lentivirus with similar results.