Encoding Thiol Metabolite Information into a DNA Sequence by Disulfide Exchange Reaction on Oligodeoxynucleotides for Parallel Analysis of the Metabolite and mRNA

Abstract

AbstractWe designed a methodology to encode the thiol metabolite information into a DNA sequence. Thiol metabolites, such as glutathione (GSH) significantly contributes to regulating redox balance in the cells. We constructed a molecular system to quantify GSH by releasing DNA from magnetic beads upon the disulfide exchange reaction. We designed the molecular system as follows: for the conversion of thiol metabolite information into DNA sequence, we used a disulfide exchange reaction between the disulfide (reaction site) tethered oligonucleotide and thiol group of target metabolites. GSH and mRNA in the cell extract were successfully quantified from the samples obtained by DNA encoding and reverse transcription. The present protocol allowed parallel analysis of thiol metabolite and mRNA.