Bisulfite-free, base-resolution analysis of 5-formylcytosine at the genome scale
Publication information:
Xia, B.; Han, D.; Lu, X.; Sun, Z.; Zhou, A.; Yin, Q.; Zeng, H.; Liu, M.; Jiang, X.; Xie, W.; He, C.; Yi, C.
Bisulfite-Free, Base-Resolution Analysis of 5-Formylcytosine at the Genome Scale. Nat Methods 2015, 12 (11), 1047-50. https://doi.org/10.1038/nmeth.3569.
Abstract
Active DNA demethylation in mammals involves oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). However, genome-wide detection of 5fC at single-base resolution remains challenging. Here we present fC-CET, a bisulfite-free method for whole-genome analysis of 5fC based on selective chemical labeling of 5fC and subsequent C-to-T transition during PCR. Base-resolution 5fC maps showed limited overlap with 5hmC, with 5fC-marked regions more active than 5hmC-marked ones.