Ligation-Free Ribosome Profiling Reveals Cell Type-Specific Regulation of Protein Synthesis in the Brain

Ribosome profiling has emerged as a powerful tool for genome-wide measurements of translation, but library construction requires multiple ligation steps and remains cumbersome relative to more conventional deep sequencing experiments. We report a new approach to ribosome profiling that does not require ligation. Library construction for ligation-free ribosome profiling can be completed in one day with as little as 1 ng of purified RNA footprints. We used ligation-free ribosome profiling to identify new patterns of cell type-specific translation in the brain and tested its ability to identify translational targets of mTOR signaling in the brain. Overall design: We report a strategy for ligation free cell-type specific ribosome profiling. Our strategy was utilized to explore cell-type specific expression in mouse brain.