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© 2021 American Chemical Society.5–Hydroxymethylcytosine (5hmC) and 5–formylcytosine (5fC) are key intermediates of active DNA demethylation, for which the global detection methods are still restricted by high cost and long operation time. Here, we demonstrate a pearl necklacelike strategy to accurately quantify global 5hmC and 5fC in genomic DNA. In this method, the metal–organic framework (MOF), [Cu3(BTC)2] (denoted as HKUST–1, H3BTC = 1,3,5–benzenetricarboxylic acid), with a diameter of ∼30 nm that contains ∼15 »000 copper ions (Cu2+) as the "super label"was in situ grown in the carboxylated 5hmC and 5fC loci of genomic DNA via the coordination between Cu2+ and the carboxyl group. After the acid digestion of MOF, the concentration of Cu2+, which has a quantitative relationship with the 5hmC/5fC content, was measured by inductively coupled plasma–atomic emission spectroscopy (ICP–AES). The metal element enrichment during MOF growth has amplified the signal by 4 orders of magnitude, realizing sensitive and accurate quantification of global 5hmC and 5fC in different tissues with a detection limit of 0.031% and 0.019‰ in DNA, respectively. The bisulfite– and mass spectrometry–free strategy is easily performed in almost all research and medical laboratories and would provide potential capability to quantify other candidate modifications in nucleotides.
