Background IL-15 plays a crucial role in regulating the function of natural killer (NK) cells, while CISH acts as a negative regulator of the IL-15 signaling pathway, thereby affecting NK cell function. Gene editing targeting the CISH and IL-15 genes in NK cells has been shown to enhance their anti-tumor activity.
Objective To investigate the effect of CISH gene and IL-15 gene on cytotoxicity of NK cells.
Methods The CISH gene was disrupted using the AsCas12a Ultra gene editing tool, while the IL-15 gene was introduced to facilitate genetic modification of NK cells. The knockout efficiency of the CISH gene was assessed through gene sequencing, and the expression of IL-15 was measured via ELISA. Subsequently, the cytotoxicity of NK cells against H358 cell line post-gene editing was evaluated using LDH assay, and the impact of CISH knockout on IL-15-mediated anti-tumor function in NK cells was examined.
Results The killing efficiency of the CISH gene knockout group and the CISH KO + IL-15 KI group was significantly enhanced compared to the control group (E : T=2 : 1 group, 34.57% ± 6.32% vs 71.40 ± 5.81% and 69.23% ± 4.28%, P < 0.001).
Conclusion The knockout of the CISH gene can enhance the IL-15 signaling pathway and augment the anti-tumor activity of NK cells.
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