Detection of CpG Methylation in G-Quadruplex Forming Sequences Using G-Quadruplex Ligands

Genomic DNA methylation is involved in many diseases and is expected to be a specific biomarker for even the pre-symptomatic diagnosis of many diseases. Thus, a rapid and inexpensive detection method is required for disease diagnosis. We have previously reported that cytosine methylation in G-quadru...

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Bibliographic Details
Published in:International journal of molecular sciences 2021-12, Vol.22 (23), p.13159
Main Authors: Hasegawa, Hijiri, Sasaki, Ikkei, Tsukakoshi, Kaori, Ma, Yue, Nagasawa, Kazuo, Numata, Shusuke, Inoue, Yuuki, Kim, Yeji, Ikebukuro, Kazunori
Format: Article
Language:English
Subjects:
Bcl-2 protein
Binding
Biomarkers
biosensor
Chemiluminescence
CpG Islands
Cytosine
Deoxyribonucleic acid
Diagnosis
DNA
DNA - chemistry
DNA - metabolism
DNA Methylation
Fluorescence
G-quadruplex
G-Quadruplexes
G4 ligand
Genomes
Humans
Ligands
Nucleotide sequence
Proto-Oncogene Proteins c-bcl-2 - chemistry
Proto-Oncogene Proteins c-bcl-2 - metabolism
Proto-Oncogene Proteins p21(ras) - chemistry
Proto-Oncogene Proteins p21(ras) - metabolism
Statistical analysis
Topology
Tumorigenesis
Vascular endothelial growth factor
Vascular Endothelial Growth Factor A - chemistry
Vascular Endothelial Growth Factor A - metabolism
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Summary:Genomic DNA methylation is involved in many diseases and is expected to be a specific biomarker for even the pre-symptomatic diagnosis of many diseases. Thus, a rapid and inexpensive detection method is required for disease diagnosis. We have previously reported that cytosine methylation in G-quadruplex (G4)-forming oligonucleotides develops different G4 topologies. In this study, we developed a method for detecting CpG methylation in G4-forming oligonucleotides based on the structural differences between methylated and unmethylated G4 DNAs. The differences in G4 topologies due to CpG methylation can be discriminated by G4 ligands. We performed a binding assay between methylated or unmethylated G4 DNAs and G4 ligands. The binding abilities of fluorescent G4 ligands to -2, , , G4-forming sequences were examined by fluorescence-based microtiter plate assay. The differences in fluorescence intensities between methylated and unmethylated G4 DNAs were statistically significant. In addition to fluorescence detection, the binding of G4 ligand to DNA was detected by chemiluminescence. A significant difference was also detected in chemiluminescence intensity between methylated and unmethylated DNA. This is the first study on the detection of CpG methylation in G4 structures, focusing on structural changes using G4 ligands.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms222313159