High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter

Abstract As one of the major epigenetic modifications, DNA methylation is constantly regulated during embryonic development, cell lineage commitment, and pathological processes. To facilitate real-time observation of DNA methylation, we generated a transgenic zebrafish reporter of DNA methylation (z...

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Autores principales:	Ranran Zhang, Lian Liu, Yuxiao Yao, Fei Fei, Feng Wang, Qian Yang, Yonghao Gui, Xu Wang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a2fa41379dbb4771a4fa4da4b49de0ab
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spelling	oai:doaj.org-article:a2fa41379dbb4771a4fa4da4b49de0ab2021-12-02T11:41:20ZHigh Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter10.1038/s41598-017-05648-82045-2322https://doaj.org/article/a2fa41379dbb4771a4fa4da4b49de0ab2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05648-8https://doaj.org/toc/2045-2322Abstract As one of the major epigenetic modifications, DNA methylation is constantly regulated during embryonic development, cell lineage commitment, and pathological processes. To facilitate real-time observation of DNA methylation, we generated a transgenic zebrafish reporter of DNA methylation (zebraRDM) via knockin of an mCherry-fused methyl-CpG binding domain (MBD) probe driven by the bactin2 promoter. The probe colocalized with heterochromatin, and its intensity was positively correlated with 5 mC immunostaining at a subcellular resolution in early embryos. Biochemical assays indicated that cells with stronger fluorescence maintained a higher level of DNA methylation, and time-lapse imaging at the blastula stage showed that the level of DNA methylation was transiently strengthened during mitosis. By crossing zebraRDM with other fluorescent transgenic lines, we demonstrate that the reporter can visually distinguish different cell lineages in organs like the heart. Our zebraRDM reporter therefore serves as a convenient and powerful tool for high-resolution investigation of methylation dynamics in live animals.Ranran ZhangLian LiuYuxiao YaoFei FeiFeng WangQian YangYonghao GuiXu WangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ranran Zhang Lian Liu Yuxiao Yao Fei Fei Feng Wang Qian Yang Yonghao Gui Xu Wang High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter
description	Abstract As one of the major epigenetic modifications, DNA methylation is constantly regulated during embryonic development, cell lineage commitment, and pathological processes. To facilitate real-time observation of DNA methylation, we generated a transgenic zebrafish reporter of DNA methylation (zebraRDM) via knockin of an mCherry-fused methyl-CpG binding domain (MBD) probe driven by the bactin2 promoter. The probe colocalized with heterochromatin, and its intensity was positively correlated with 5 mC immunostaining at a subcellular resolution in early embryos. Biochemical assays indicated that cells with stronger fluorescence maintained a higher level of DNA methylation, and time-lapse imaging at the blastula stage showed that the level of DNA methylation was transiently strengthened during mitosis. By crossing zebraRDM with other fluorescent transgenic lines, we demonstrate that the reporter can visually distinguish different cell lineages in organs like the heart. Our zebraRDM reporter therefore serves as a convenient and powerful tool for high-resolution investigation of methylation dynamics in live animals.
format	article
author	Ranran Zhang Lian Liu Yuxiao Yao Fei Fei Feng Wang Qian Yang Yonghao Gui Xu Wang
author_facet	Ranran Zhang Lian Liu Yuxiao Yao Fei Fei Feng Wang Qian Yang Yonghao Gui Xu Wang
author_sort	Ranran Zhang
title	High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter
title_short	High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter
title_full	High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter
title_fullStr	High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter
title_full_unstemmed	High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter
title_sort	high resolution imaging of dna methylation dynamics using a zebrafish reporter
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/a2fa41379dbb4771a4fa4da4b49de0ab
work_keys_str_mv	AT ranranzhang highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT lianliu highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT yuxiaoyao highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT feifei highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT fengwang highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT qianyang highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT yonghaogui highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter AT xuwang highresolutionimagingofdnamethylationdynamicsusingazebrafishreporter
_version_	1718395416443092992

High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter

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