Knockout of AMD-associated Gene Reduces Mitochondrial Superoxide in Human Retinal Pigment Epithelial Cells

Genetic and epidemiologic studies have significantly advanced our understanding of the genetic factors contributing to age-related macular degeneration (AMD). In particular, recent expression quantitative trait loci (eQTL) studies have highlighted as a significant gene that confers risk of developing AMD. However, the role of in retinal cells such as retinal pigment epithelium (RPE) and how it contributes to AMD pathology are unknown. Here we report the generation of a stable human RPE cell line ARPE-19 with knockout using CRISPR/Cas, providing an model to investigate the functions of . We conducted functional studies on the knockout cell line and showed that it retained normal levels of cell proliferation, cell viability, phagocytosis and autophagy. Also, we performed RNA sequencing to profile the transcriptome of knockout cells. Our results highlighted significant changes in genes involved in immune response, complement activation, oxidative damage and vascular development. We showed that loss of caused a reduction in mitochondrial superoxide levels, which is consistent with the upregulation of the mitochondrial superoxide dismutase . In conclusion, this study demonstrates a novel link between and in ARPE-19, which supports a potential role of in regulating oxidative stress in AMD pathology.