title: Ligation-free ribosome profiling of cell type-specific translation in the brain.
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.
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, NY, 10032, USA.|Columbia University M.D./Ph.D. Program, Columbia University Medical Center, New York, NY, 10032, USA.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Hornstein
authors.affiliation: Graduate Ph.D. Program in Pharmacology and Molecular Signaling, Columbia University Medical Center, New York, NY, 10032, USA.|Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, 10032, USA.
authors.forename: Daniela
authors.initials: D
authors.lastname: Torres
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, NY, 10032, USA.
authors.forename: Sohani
authors.initials: S
authors.lastname: Das Sharma
authors.affiliation: Department of Neurology, Columbia University Medical Center, New York, NY, 10032, USA.
authors.forename: Guomei
authors.initials: G
authors.lastname: Tang
authors.affiliation: Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, 10032, USA. pc561@cumc.columbia.edu.
authors.forename: Peter
authors.initials: P
authors.lastname: Canoll
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, NY, 10032, USA. pas2182@columbia.edu.|Department of Biochemistry and Molecular Biophysics, Columbia University Medical Center, New York, NY, 10032, USA. pas2182@columbia.edu.|Columbia Sulzberger Genome Center, Columbia University Medical Center, New York, NY, 10032, USA. pas2182@columbia.edu.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Sims
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D051379:Mice; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000235:genetics*
chemical_list: D012313:RNA; C546843:mTOR protein, mouse; D058570:TOR Serine-Threonine Kinases
keywords: Brain; Ribosome profiling; Translation