PUBLICATION

High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter

Authors
Zhang, R., Liu, L., Yao, Y., Fei, F., Wang, F., Yang, Q., Gui, Y., Wang, X.
ID
ZDB-PUB-170716-5
Date
2017
Source
Scientific Reports   7: 5430 (Journal)
Registered Authors
Wang, Xu
Keywords
DNA methylation, Zebrafish
MeSH Terms
  • Genes, Reporter/genetics*
  • Animals
  • Microscopy, Confocal
  • Embryo, Nonmammalian/diagnostic imaging
  • Embryo, Nonmammalian/embryology
  • Embryo, Nonmammalian/metabolism*
  • DNA Methylation*
  • Promoter Regions, Genetic/genetics
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Animals, Genetically Modified
  • Kinetics
  • Heterochromatin/genetics
  • Heterochromatin/metabolism
  • Luminescent Proteins/genetics
  • Luminescent Proteins/metabolism
(all 17)
PubMed
28710355 Full text @ Sci. Rep.
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.
Genes / Markers
Figures
Figure Gallery (7 images)
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Expression
No data available
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
f1TgTransgenic Insertion
    fdu6TgTransgenic Insertion
      ia4TgTransgenic Insertion
        1 - 3 of 3
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        Human Disease / Model
        No data available
        Sequence Targeting Reagents
        No data available
        Fish
        No data available
        Antibodies
        No data available
        Orthology
        No data available
        Engineered Foreign Genes
        Marker Marker Type Name
        EGFPEFGEGFP
        GFPEFGGFP
        mCherryEFGmCherry
        1 - 3 of 3
        Show
        Mapping
        No data available
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        Citation
        Zhang, R., Liu, L., Yao, Y., Fei, F., Wang, F., Yang, Q., Gui, Y., Wang, X. (2017) High Resolution Imaging of DNA Methylation Dynamics using a Zebrafish Reporter. Scientific Reports. 7:5430.