A genome-scale ORF screen for resilience factors that protect the RPE and restore vision in aged mice.

Aging is a complex process, yet therapeutic discovery has largely relied on hypothesis-driven testing of individual targets. Here, we performed an unbiased genome-scale open reading frame (ORF) screen for oxidative-stress resistance in retinal pigment epithelium (RPE) cells, which are particularly vulnerable to oxidative damage with age. The screen identified nine protective ORFs, including antioxidant factors and a regulator of epithelial-to-mesenchymal transition. Notably, we also identified three factors that reduce transcriptomic age, including the cardiac transcription factor NKX2-5. NKX2-5 conferred robust oxidative resilience in vitro and in vivo . Domain dissection showed that its homeodomain was dispensable for resilience, enabling development of NKX2-5ΔHD, which retained protective activity without detectable toxicity over 14 months. In middle-aged mice, subretinal NKX2-5ΔHD restored visual and electrophysiological function, and its systemic delivery improved grip strength and reduced frailty in late-aged mice. These findings establish genome-scale ORF screening as a strategy to discover transferable resilience factors that can be reengineered to counter age-related functional decline across tissues.