Sequence-specific microscopic visualization of DNA methylation status at satellite repeats in individual cell nuclei and chromosomes

Yufeng Li, Yusuke Miyanari, Kenjiro Shirane, Hirohisa Nitta, Takeo Kubota, Hirofumi Ohashi, Akimitsu Okamoto, Hiroyuki Sasaki

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)e186
JournalNucleic acids research
Volume41
Issue number19
DOIs
Publication statusPublished - Oct 2013

All Science Journal Classification (ASJC) codes

  • Genetics

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