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

Lu, Y. R. R.; Shen, H.; Kajderowicz, K.; Ai, J.; Chen-Neumann, Y. H.; Hu, Y.; Tyshkovskiy, A.; Lou, Q.; Dong, T.; Grauvogel, L.; Chen, Z.; Cameron, J. C.; Li, X.; Shirahama, S.; Bligh, M.; Hou, S.; Chin, J.; Costa, M. W.; Ryu, J.; Sharma, R.; Mann, M.; Bharti, K.; Bowes Rickman, C.; Gladyshev, V. N.; Gregory-Ksander, M.; Ksander, B.; Weissman, J.

doi:10.64898/2026.09.29.755187

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bioRxiv · 10.64898/2026.09.29.755187

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

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Abstract

Aging is a complex phenomenon, yet therapeutic discovery has largely relied on testing individual hypothesis-driven targets. Here, we performed a genome-scale open reading frame (ORF) screen for oxidative-stress resistance in retinal pigment epithelium (RPE) cells, which are vulnerable to oxidative damage during aging. The screen identified nine protective ORFs including antioxidant factors and an epithelial-to-mesenchymal transition regulator. 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 revealed that its homeodomain is dispensable for resilience, enabling development of NKX2-5{Delta}HD, which retained protective activity without detectable toxicity over 14 months. In middle-aged mice, subretinal NKX2-5{Delta}HD restored visual and electrophysiological function, and its systemic delivery improved grip strength and reduced frailty in late-aged mice. These findings highlight the value of unbiased, genome-scale gain-of-function screening, as resilience factors normally expressed in other contexts can be reengineered as potential therapeutics for age-related decline in other tissues.

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Lu, Y. R. R., Shen, H., Kajderowicz, K., Ai, J., Chen-Neumann, Y. H., Hu, Y., Tyshkovskiy, A., Lou, Q., Dong, T., Grauvogel, L., Chen, Z., Cameron, J. C., Li, X., Shirahama, S., Bligh, M., Hou, S., Chin, J., Costa, M. W., Ryu, J., Sharma, R., Mann, M., Bharti, K., Bowes Rickman, C., Gladyshev, V. N., Gregory-Ksander, M., Ksander, B., Weissman, J.. 2026-09-30. A genome-scale ORF screen for resilience factors that protect the RPE and restore vision in aged mice. https://doi.org/10.64898/2026.09.29.755187

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