Since MASLD occurs as a mitochondrial disease and the PNPLA3, MBOAT7 and TM6SF2 loss-of-function mutations seem to achieve a role in organelles’ dysfunction, in this study, we firstly investigated the impact of gene deletion on mitochondrial dynamism and integrity in KO HepG2 cells. This evidence concerns the gene PNPLA3 and metabolic dysfunction-associated steatotic liver disease.