Ligation-free ribosome profiling of cell type-specific translation in the brain

Genome Biol. 2016 Jul 5;17(1):149. doi: 10.1186/s13059-016-1005-1.

Abstract

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, ligation-free 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 apply ligation-free ribosome profiling to mouse brain tissue to identify new patterns of cell type-specific translation and test its ability to identify translational targets of mTOR signaling in the brain.

Keywords: Brain; Ribosome profiling; Translation.

MeSH terms

  • Animals
  • Brain / metabolism
  • High-Throughput Nucleotide Sequencing*
  • Mice
  • Protein Biosynthesis
  • RNA / genetics*
  • RNA / metabolism
  • Ribosomes / genetics*
  • Ribosomes / metabolism
  • Signal Transduction
  • TOR Serine-Threonine Kinases / genetics*

Substances

  • RNA
  • mTOR protein, mouse
  • TOR Serine-Threonine Kinases