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Ligation-free ribosome profiling of cell type-specific translation 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, ligation-free approach to ribosome profiling tha...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4934013/ https://www.ncbi.nlm.nih.gov/pubmed/27380875 http://dx.doi.org/10.1186/s13059-016-1005-1 |
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author | Hornstein, Nicholas Torres, Daniela Das Sharma, Sohani Tang, Guomei Canoll, Peter Sims, Peter A. |
author_facet | Hornstein, Nicholas Torres, Daniela Das Sharma, Sohani Tang, Guomei Canoll, Peter Sims, Peter A. |
author_sort | Hornstein, Nicholas |
collection | PubMed |
description | 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13059-016-1005-1) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4934013 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-49340132016-07-07 Ligation-free ribosome profiling of cell type-specific translation in the brain Hornstein, Nicholas Torres, Daniela Das Sharma, Sohani Tang, Guomei Canoll, Peter Sims, Peter A. Genome Biol Method 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13059-016-1005-1) contains supplementary material, which is available to authorized users. BioMed Central 2016-07-05 /pmc/articles/PMC4934013/ /pubmed/27380875 http://dx.doi.org/10.1186/s13059-016-1005-1 Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Method Hornstein, Nicholas Torres, Daniela Das Sharma, Sohani Tang, Guomei Canoll, Peter Sims, Peter A. Ligation-free ribosome profiling of cell type-specific translation in the brain |
title | Ligation-free ribosome profiling of cell type-specific translation in the brain |
title_full | Ligation-free ribosome profiling of cell type-specific translation in the brain |
title_fullStr | Ligation-free ribosome profiling of cell type-specific translation in the brain |
title_full_unstemmed | Ligation-free ribosome profiling of cell type-specific translation in the brain |
title_short | Ligation-free ribosome profiling of cell type-specific translation in the brain |
title_sort | ligation-free ribosome profiling of cell type-specific translation in the brain |
topic | Method |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4934013/ https://www.ncbi.nlm.nih.gov/pubmed/27380875 http://dx.doi.org/10.1186/s13059-016-1005-1 |
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