RRC ID |
44619
|
著者 |
Li Y, Miyanari Y, Shirane K, Nitta H, Kubota T, Ohashi H, Okamoto A, Sasaki H.
|
タイトル |
Sequence-specific microscopic visualization of DNA methylation status at satellite repeats in individual cell nuclei and chromosomes.
|
ジャーナル |
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.
|
巻・号 |
41(19)
|
ページ |
e186
|
公開日 |
2013-10-1
|
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 |
11.502
|
引用数 |
19
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
ヒト・動物細胞 |
HEV0190 |