In vivo single-cell ribosome profiling reveals cell-type-specific translational programs during aging

Yet, how aging reshapes the translational landscape of stem cells and how these changes impact their regenerative capacity remains poorly understood. Here, we present an in vivo single-cell ribosome profiling strategy to monitor tissue-wide translational landscapes of the young and aged mouse skin.

By implementing efficient in vivo cell isolation and switching to RNAse I-based ribonuclease for generating ribosomal footprints, we enhance the accessibility of single-cell ribosome profiling and facilitate the evaluation of triplet periodicity, a hallmark of high-quality data. Leveraging this strategy and integrating single-cell RNA sequencing with ribosome profiling, we document the translational landscapes of the major epidermal cell types, outline cell-type-specific translational efficiencies and capture heterogeneity in differentiation commitment among stem cell populations.