In vivo single-cell ribosome profiling reveals cell-type-specific translational programs during aging [scRNA-Seq]

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.