Hasegawa, Hijiri LG Japan Lab
Sasaki, Ikkei Tokyo University of Agriculture and Technology
Tsukakoshi, Kaori Tokyo University of Agriculture and Technology
Ma, Yue Tokyo University of Agriculture and Technology
Nagasawa, Kazuo Tokyo University of Agriculture and Technology
Numata, Shusuke Tokushima University Tokushima University Educator and Researcher Directory KAKEN Search Researchers
Inoue, Yuuki LG Japan Lab
Kim, Yeji LG Japan Lab
Ikebukuro, Kazunori Tokyo University of Agriculture and Technology
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 BCL-2, HRAS1, HRAS2, VEGF 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.
International Journal of Molecular Sciences
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
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ijms_22_23_13159.pdf 1.63 MB