Pyrene‐Based Quantitative Detection of the 5‐Formylcytosine Loci Symmetry in the CpG Duplex Content during TET‐Dependent Demethylation |
| |
| Authors: | Dr Liang Xu Ying‐Chu Chen Jenny Chong Dr Andrea Fin Dr Lisa S McCoy Dr Jun Xu Prof?Dr Chao Zhang Prof?Dr Dong Wang |
| |
| Institution: | 1. Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA 92093 (USA);2. Department of Chemistry, University of Southern California, Los Angeles, CA 90089 (USA);3. Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093 (USA) |
| |
| Abstract: | Methylcytosine (5mC) is mostly symmetrically distributed in CpG sites. Ten‐eleven‐translocation (TET) proteins are the key enzymes involved in active DNA demethylation through stepwise oxidation of 5mC. However, oxidation pathways of TET enzymes in the symmetrically methylated CpG context are still elusive. Employing the unique fluorescence properties of pyrene group, we designed and synthesized a sensitive fluorescence‐based probe not only to target 5‐formylcytosine (5fC) sites, but also to distinguish symmetric from asymmetric 5fC sites in the double stranded DNA context during TET‐dependent 5mC oxidation process. Using this novel probe, we revealed dominant levels of symmetric 5fC among total 5fC sites during in vitro TET‐dependent 5mC oxidation and novel mechanistic insights into the TET‐dependent 5mC oxidation in the mCpG context. |
| |
| Keywords: | 5‐formylcytosine DNA demethylation pyrene fluorescence TET oxidation |
|
|
