Cytosine methylation is a dynamic form of epigenetic regulation, and demethylation occurs via intermediates such as 5-formylcytosine (5-fC) that may themselves play regulatory roles. The position of methylated cytosines (5-mC) in the genome can be inferred using bisulfite to convert unmodified cytosine to thymine during sequencing (5-fC is also read as thymine in the process). Booth et al. have developed reduced bisulfite sequencing (redBS-Seq), which uses sodium borohydride to specifically reduce 5-fC and thereby protect it from bisulfite conversion. redBS-Seq provides single-base resolution and may be more efficient than the related fCABs-Seq (chemically assisted bisulfite sequencing) method, making the genome-wide discovery of this rare mark potentially more practical. Booth et al. sequenced 5-fC in addition to 5-mC and another intermediate in mouse embryonic stem cell genomes.
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5-formylcytosine-seq at single-base resolution. Nat Methods 11, 475 (2014). https://doi.org/10.1038/nmeth.2944
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DOI: https://doi.org/10.1038/nmeth.2944