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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...

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Autores principales: Lukoseviciute, Martyna, Ling, Irving T.C., Senanayake, Upeka, Candido-Ferreira, Ivan, Taylor, Gunes, Williams, Ruth M., Sauka-Spengler, Tatjana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
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).
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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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