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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...

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Detalles Bibliográficos
Autores principales: Fujita, Toshitsugu, Asano, Yoshinori, Ohtsuka, Junko, Takada, Yoko, Saito, Kazunobu, Ohki, Rieko, Fujii, Hodaka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2013
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.
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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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