To investigate whether NECTIN2 expressed on HCC cells would lead to T cell exhaustion, we established Nectin2 knockout (KO) stable cells in a mouse HCC cell line, Hepa1–6, with the CRISPR–Cas9 KO system using two independent single-guide RNAs (sgRNAs). The gene discussed is NECTIN2; the disease is hepatocellular carcinoma.