biostudies-literatureUnknown6(1)Huayta JIn many organisms, dietary restriction (DR) leads to lifespan extension through the activation of cell protection and pro-longevity gene expression programs. In the nematode C. elegans, the DAF-16 transcription factor is a key aging regulator that governs the Insulin/IGF-1 signaling pathway and undergoes translocation from the cytoplasm to the nucleus of cells when animals are exposed to food limitation. However, how large is the influence of DR on DAF-16 activity, and its subsequent impact on lifespan has not been quantitatively determined. In this work, we assess the endogenous activity of DAF-16 under various DR regimes by coupling CRISPR/Cas9-enabled fluorescent tagging of DAF-16 with quantitative image analysis and machine learning. Our results indicate that DR regimes induce strong eCommunications biology203https://www.ebi.ac.uk/biostudies/studies/S-EPMC9941123biostudies-literatureEndogenous DAF-16 spatiotemporal activity quantitatively predicts lifespan extension induced by dietary restriction.PMC9941123Crapster JPSan-Miguel AHuayta JfalseEndogenous DAF-16 spatiotemporal activity quantitatively predicts lifespan extension induced by dietary restriction.In many organisms, dietary restriction (DR) leads to lifespan extension through the activation of cell protection and pro-longevity gene expression programs. In the nematode C. elegans, the DAF-16 transcription factor is a key aging regulator that governs the Insulin/IGF-1 signaling pathway and undergoes translocation from the cytoplasm to the nucleus of cells when animals are exposed to food limitation. However, how large is the influence of DR on DAF-16 activity, and its subsequent impact on lifespan has not been quantitatively determined. In this work, we assess the endogenous activity of DAF-16 under various DR regimes by coupling CRISPR/Cas9-enabled fluorescent tagging of DAF-16 with quantitative image analysis and machine learning. Our results indicate that DR regimes induce strong e2023-01-01T00:00:00Z2023 Feb2025-04-04T13:36:23.217Z2025-02-18T22:55:47.034ZS-EPMC99411233680764610.1038/s42003-023-04562-2