RRC ID 44619
Author Li Y, Miyanari Y, Shirane K, Nitta H, Kubota T, Ohashi H, Okamoto A, Sasaki H.
Title Sequence-specific microscopic visualization of DNA methylation status at satellite repeats in individual cell nuclei and chromosomes.
Journal Nucleic Acids Res.
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.
Volume 41(19)
Pages e186
Published 2013-10
DOI 10.1093/nar/gkt766
PII gkt766
PMID 23990328
PMC PMC3799461
MeSH 5-Methylcytosine / analogs & derivatives Animals Base Sequence Cell Nucleus / genetics* Cells, Cultured Chromosomes, Mammalian / chemistry* Cytosine / analogs & derivatives Cytosine / analysis DNA Methylation* DNA, Satellite* Embryonic Stem Cells / chemistry Humans In Situ Hybridization, Fluorescence / methods* Male Mice Spermatozoa / chemistry
IF 11.561
Times Cited 11
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells