Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay

We describe a fluorescence recovery after photobleaching (FRAP) protocol for assessing the dynamics of heterochromatin/euchromatin and identifying chromatin relaxers for cell fate transition. Here, we developed a system to track heterochromatin foci with HP1α-cherry and performed FRAP assay of H1-GF...

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Autores principales: Long, Qi, Qi, Juntao, Li, Wei, Zhou, Yanshuang, Chen, Keshi, Wu, Hao, Liu, Xingguo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8348307/
https://www.ncbi.nlm.nih.gov/pubmed/34401775
http://dx.doi.org/10.1016/j.xpro.2021.100706
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author Long, Qi
Qi, Juntao
Li, Wei
Zhou, Yanshuang
Chen, Keshi
Wu, Hao
Liu, Xingguo
author_facet Long, Qi
Qi, Juntao
Li, Wei
Zhou, Yanshuang
Chen, Keshi
Wu, Hao
Liu, Xingguo
author_sort Long, Qi
collection PubMed
description We describe a fluorescence recovery after photobleaching (FRAP) protocol for assessing the dynamics of heterochromatin/euchromatin and identifying chromatin relaxers for cell fate transition. Here, we developed a system to track heterochromatin foci with HP1α-cherry and performed FRAP assay of H1-GFP to analyze the dynamics of heterochromatin and euchromatin during somatic cell reprogramming. This protocol is used to screen factors that impact chromatin structure, which could also be used to identify chromatin relaxers and repressors in various cell fate transitions. For complete details on the use and execution of this protocol, please refer to Chen et al. (2016) and Chen et al. (2020).
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spelling pubmed-83483072021-08-15 Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay Long, Qi Qi, Juntao Li, Wei Zhou, Yanshuang Chen, Keshi Wu, Hao Liu, Xingguo STAR Protoc Protocol We describe a fluorescence recovery after photobleaching (FRAP) protocol for assessing the dynamics of heterochromatin/euchromatin and identifying chromatin relaxers for cell fate transition. Here, we developed a system to track heterochromatin foci with HP1α-cherry and performed FRAP assay of H1-GFP to analyze the dynamics of heterochromatin and euchromatin during somatic cell reprogramming. This protocol is used to screen factors that impact chromatin structure, which could also be used to identify chromatin relaxers and repressors in various cell fate transitions. For complete details on the use and execution of this protocol, please refer to Chen et al. (2016) and Chen et al. (2020). Elsevier 2021-07-30 /pmc/articles/PMC8348307/ /pubmed/34401775 http://dx.doi.org/10.1016/j.xpro.2021.100706 Text en © 2021 The Author(s) https://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
Long, Qi
Qi, Juntao
Li, Wei
Zhou, Yanshuang
Chen, Keshi
Wu, Hao
Liu, Xingguo
Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay
title Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay
title_full Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay
title_fullStr Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay
title_full_unstemmed Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay
title_short Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay
title_sort protocol for detecting chromatin dynamics and screening chromatin relaxer by frap assay
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8348307/
https://www.ncbi.nlm.nih.gov/pubmed/34401775
http://dx.doi.org/10.1016/j.xpro.2021.100706
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