To assess the efficacy with which NKG2D-CAR T cells can target NKG2DL-expressing cells, we first established, using the PiggyBac transposon system, B16F10 melanoma cells individually overexpressing one of three classes of mouse NKG2DLs: histocompatibility antigen 60a (H60a), retinoic acid early transcript 1β (Rae1β) or murine UL16-binding-protein-like transcript 1 (Mult1) (Fig. 2A). This evidence concerns the gene KLRK1 and melanoma.