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Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP)
Biochemical analysis of molecular interactions in specific genomic regions requires their isolation while retaining molecular interactions in vivo. Here, we report isolation of telomeres by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using a transcription activato...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3821016/ https://www.ncbi.nlm.nih.gov/pubmed/24201379 http://dx.doi.org/10.1038/srep03171 |
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author | Fujita, Toshitsugu Asano, Yoshinori Ohtsuka, Junko Takada, Yoko Saito, Kazunobu Ohki, Rieko Fujii, Hodaka |
author_facet | Fujita, Toshitsugu Asano, Yoshinori Ohtsuka, Junko Takada, Yoko Saito, Kazunobu Ohki, Rieko Fujii, Hodaka |
author_sort | Fujita, Toshitsugu |
collection | PubMed |
description | Biochemical analysis of molecular interactions in specific genomic regions requires their isolation while retaining molecular interactions in vivo. Here, we report isolation of telomeres by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using a transcription activator-like (TAL) protein recognizing telomere repeats. Telomeres recognized by the tagged TAL protein were immunoprecipitated with an antibody against the tag and subjected to identification of telomere-binding molecules. enChIP-mass spectrometry (enChIP-MS) targeting telomeres identified known and novel telomere-binding proteins. The data have been deposited to the ProteomeXchange with identifier PXD000461. In addition, we showed that RNA associated with telomeres could be isolated by enChIP. Identified telomere-binding molecules may play important roles in telomere biology. enChIP using TAL proteins would be a useful tool for biochemical analysis of specific genomic regions of interest. |
format | Online Article Text |
id | pubmed-3821016 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-38210162013-11-09 Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) Fujita, Toshitsugu Asano, Yoshinori Ohtsuka, Junko Takada, Yoko Saito, Kazunobu Ohki, Rieko Fujii, Hodaka Sci Rep Article Biochemical analysis of molecular interactions in specific genomic regions requires their isolation while retaining molecular interactions in vivo. Here, we report isolation of telomeres by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using a transcription activator-like (TAL) protein recognizing telomere repeats. Telomeres recognized by the tagged TAL protein were immunoprecipitated with an antibody against the tag and subjected to identification of telomere-binding molecules. enChIP-mass spectrometry (enChIP-MS) targeting telomeres identified known and novel telomere-binding proteins. The data have been deposited to the ProteomeXchange with identifier PXD000461. In addition, we showed that RNA associated with telomeres could be isolated by enChIP. Identified telomere-binding molecules may play important roles in telomere biology. enChIP using TAL proteins would be a useful tool for biochemical analysis of specific genomic regions of interest. Nature Publishing Group 2013-11-08 /pmc/articles/PMC3821016/ /pubmed/24201379 http://dx.doi.org/10.1038/srep03171 Text en Copyright © 2013, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/3.0/ This work is licensed under a Creative Commons Attribution 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by/3.0/ |
spellingShingle | Article Fujita, Toshitsugu Asano, Yoshinori Ohtsuka, Junko Takada, Yoko Saito, Kazunobu Ohki, Rieko Fujii, Hodaka Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) |
title | Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) |
title_full | Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) |
title_fullStr | Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) |
title_full_unstemmed | Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) |
title_short | Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) |
title_sort | identification of telomere-associated molecules by engineered dna-binding molecule-mediated chromatin immunoprecipitation (enchip) |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3821016/ https://www.ncbi.nlm.nih.gov/pubmed/24201379 http://dx.doi.org/10.1038/srep03171 |
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