DNA中の酸化損傷塩基を認識し検出を可能とする人工核酸の創製研究

抄録

To determine of the position of oxidative nucleoside in DNA, the selective detection of 8-oxo-2’-deoxyguanosine (8-oxo-dG) without chemical or enzymatic treatment is an attractive tool for genomic research. Here, I described the summary of our resent study which designed and synthesized the non-natural nucleoside analogue, the adenosine-1,3-diazaphenoxazine derivative (Adap), for selective recognition of 8-oxo-dG in DNA. This study has clearly shown that Adap has stabilizing effect of the duplex including the Adap-8-oxodG base pair with high selectivity. Furthermore, the fluorescent quenching property of Adap has been shown to detect of 8-oxo-dG in DNA. And also, the triphosphate of Adap (dAdapTP) was synthesized and tested for single nucleotide incorporation reaction to primer strand using DNA polymerase. The efficiency of dAdapTP incorporation into 8-oxo-dG-containing templates was better than with dG-containing templates. Then the detection of 8-oxo-dG in human telomeric DNA sequences extracted from H₂O₂-treated HeLa cells and in genome DNA extracted from small intestine of transgenic mice were successful. Furthermore, I will introduce the development of Adap derivatives, which are expected to be applied to sequencing technology in this review.