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Updated: May 8, 2026
DNA Methylation: Bisulphite Modification and Analysis
Published on: October 21, 2011
Sequence-specific microscopic visualization of DNA methylation status at satellite repeats in individual cell nuclei
Yufeng Li1, Yusuke Miyanari, Kenjiro Shirane
1Division of Epigenomics and Development, Medical Institute of Bioregulation, and Epigenome Network Research Center, Kyushu University, Fukuoka 812-8582, Japan, The Cancer Institute, Tangshan People's Hospital, Hebei 063001, China, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Université de Strasbourg, F-67404 Illkirch, Cité Universitaire de Strasbourg, France, Department of Epigenetics Medicine, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Yamanashi 409-3898, Japan, Division of Medical Genetics, Saitama Children's Medical Center, Saitama 339-8551, Japan and Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo 113-8656, Japan.
Abstract:
Methylation-specific fluorescence in situ hybridization (MeFISH) was developed for microscopic visualization of DNA methylation status at specific repeat sequences in individual cells. MeFISH is based on the differential reactivity of 5-methylcytosine and cytosine in target DNA for interstrand complex formation with osmium and bipyridine-containing nucleic acids (ICON). Cell nuclei and chromosomes hybridized with fluorescence-labeled ICON probes for mouse major and minor satellite repeats were treated with osmium for crosslinking. After denaturation, fluorescent signals were retained specifically at satellite repeats in wild-type, but not in DNA methyltransferase triple-knockout (negative control) mouse embryonic stem cells. Moreover, using MeFISH, we successfully detected hypomethylated satellite repeats in cells from patients with immunodeficiency, centromeric instability and facial anomalies syndrome and 5-hydroxymethylated satellite repeats in male germ cells, the latter of which had been considered to be unmethylated based on anti-5-methylcytosine antibody staining. MeFISH will be suitable for a wide range of applications in epigenetics research and medical diagnosis.
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