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Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken
Chromatin immunoprecipitation with sequencing (ChIP-seq) has been instrumental in understanding transcription factor (TF) binding during gene regulation. ChIP-seq requires specific antibodies against desired TFs, which are not available for numerous species. Here, we describe a tissue-specific bioti...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580215/ https://www.ncbi.nlm.nih.gov/pubmed/33111104 http://dx.doi.org/10.1016/j.xpro.2020.100066 |
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author | Lukoseviciute, Martyna Ling, Irving T.C. Senanayake, Upeka Candido-Ferreira, Ivan Taylor, Gunes Williams, Ruth M. Sauka-Spengler, Tatjana |
author_facet | Lukoseviciute, Martyna Ling, Irving T.C. Senanayake, Upeka Candido-Ferreira, Ivan Taylor, Gunes Williams, Ruth M. Sauka-Spengler, Tatjana |
author_sort | Lukoseviciute, Martyna |
collection | PubMed |
description | Chromatin immunoprecipitation with sequencing (ChIP-seq) has been instrumental in understanding transcription factor (TF) binding during gene regulation. ChIP-seq requires specific antibodies against desired TFs, which are not available for numerous species. Here, we describe a tissue-specific biotin ChIP-seq protocol for zebrafish and chicken embryos which utilizes AVI tagging of TFs, permitting their biotinylation by a co-expressed nuclear biotin ligase. Subsequently, biotinylated factors can be precipitated with streptavidin beads, enabling the user to construct TF genome-wide binding landscapes like conventional ChIP-seq methods. For complete details on the use and execution of this protocol, please see Lukoseviciute et al. (2018) and Ling and Sauka-Spengler (2019). |
format | Online Article Text |
id | pubmed-7580215 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-75802152020-10-26 Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken Lukoseviciute, Martyna Ling, Irving T.C. Senanayake, Upeka Candido-Ferreira, Ivan Taylor, Gunes Williams, Ruth M. Sauka-Spengler, Tatjana STAR Protoc Protocol Chromatin immunoprecipitation with sequencing (ChIP-seq) has been instrumental in understanding transcription factor (TF) binding during gene regulation. ChIP-seq requires specific antibodies against desired TFs, which are not available for numerous species. Here, we describe a tissue-specific biotin ChIP-seq protocol for zebrafish and chicken embryos which utilizes AVI tagging of TFs, permitting their biotinylation by a co-expressed nuclear biotin ligase. Subsequently, biotinylated factors can be precipitated with streptavidin beads, enabling the user to construct TF genome-wide binding landscapes like conventional ChIP-seq methods. For complete details on the use and execution of this protocol, please see Lukoseviciute et al. (2018) and Ling and Sauka-Spengler (2019). Elsevier 2020-07-31 /pmc/articles/PMC7580215/ /pubmed/33111104 http://dx.doi.org/10.1016/j.xpro.2020.100066 Text en © 2020. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Lukoseviciute, Martyna Ling, Irving T.C. Senanayake, Upeka Candido-Ferreira, Ivan Taylor, Gunes Williams, Ruth M. Sauka-Spengler, Tatjana Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken |
title | Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken |
title_full | Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken |
title_fullStr | Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken |
title_full_unstemmed | Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken |
title_short | Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken |
title_sort | tissue-specific in vivo biotin chromatin immunoprecipitation with sequencing in zebrafish and chicken |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580215/ https://www.ncbi.nlm.nih.gov/pubmed/33111104 http://dx.doi.org/10.1016/j.xpro.2020.100066 |
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