Sequence-selective 5-methylcytosine oxidation for epigenotyping.
Methylation of DNA is an epigenetic modification that can play an important role in the control of gene expression in mammalian cells. The development of a simple and convenient method for site-specific discrimination of cytosine methylation is imperative for genomic studies. Here we report a facile method for distinguishing between cytosine and 5-methylcytosine. Osmium tetroxide caused the dihydroxylation of the C5-C6 double bond of 5-methylcytosine under an appropriate reaction conditions. The oxidation of 5-methylcytosine-containing target DNA was controlled by hybridization with a guide DNA. This technique facilitates the typing of cytosine methylation at a specific site of the target DNA.
Okamoto
Akimitsu
A
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Kyoto 615-8510, Japan.
Tainaka
Kazuki
K
D044503:5-Methylcytosine / Q000032:analysis* / Q000737:chemistry; D001483:Base Sequence; D003596:Cytosine / Q000032:analysis; D004247:DNA / Q000737:chemistry; D019175:DNA Methylation; D005838:Genotype; D006900:Hydroxylation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009993:Osmium Tetroxide / Q000737:chemistry; D010084:Oxidation-Reduction
D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D009993:Osmium Tetroxide