Direct, bisulfite-free 5mC and 5hmC sequencing at single-cell resolution
Xiufei CheniD, Jingfei ChengiD, Linzhen Kong, Xiao Shu, Haiqi XuiD, Masato Inoue, Marion Silvana Fernández-BerrocaliD, Dagny Sanden Døskeland, Magnar BjøråsiD, Shivan SivakumariD, Yibin LiuiD, Jing YeiD, Chun‐Xiao SongiD
Abstract
We report the first direct sequencing methodologies for quantitative detection of 5mC and 5hmC at single-base resolution and single-cell level, termed scTAPS (for 5mC + 5hmC) and scCAPS+ (for 5hmC specifically). With [~]90% mapping efficiency, our methods accurately benchmark 5mC and 5hmC profiles in CD8+ T and mES cells, respectively. Notably, scCAPS+ revealed a global increase in 5hmC within the hippocampus of aging mice, both in neurons and in non-neurons.
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