Epigenetics
Can you map DNA methylation in one cell?
Open access · cc by · source: Europe PMC
An optimized bisulfite protocol produced 82 single-cell methylomes (median ~1.4 million CpGs each); pooling a few dozen cells covered >90% of CpGs and a region-set method placed hematopoietic and ESC samples on treatment trajectories.
Study at a glance
- Design
- Animal / in-vitro — Optimized μWGBS/scWGBS plus a region-set bioinformatic method applied to K562 azacytidine, HL60 vitamin D3, and mouse ESC 2i/ATRA/EB models
- N
- N=82 · 82 single-cell methylomes, 89 four-cell methylomes, and 53 optimization/QC libraries after >80% QC pass
- Population
- Human K562, HL60, and KBM7 lines plus mouse CCE ESCs (and 32D clonal-expansion tests)
- Outcome
- Single-cell methylome coverage, clustering by cell state, and inferred epigenomic dynamics from sparse data
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Key findings
>80% of libraries passed QC (82 one-cell, 89 four-cell). Human one-cell: median 4.6 million reads and 1.4 million CpGs. Combining a few dozen cells covered >90% of CpGs. K562 showed global demethylation after azacytidine; HL60 changes were subtler; ESC 2i vs serum residuals placed ATRA/EB samples in the opposite direction.
Methodology
FACS-sorted 1–500 cells, ran post-bisulfite adaptor WGBS (μWGBS/scWGBS), and aggregated methylation over public region sets. Models: K562 + azacytidine, HL60 + vitamin D3, and mouse ESCs in 2i vs ATRA/embryoid-body differentiation.
Limitations
Low-coverage single-cell WGBS cannot call every CpG in one cell; cell-line models are not embryos or tumors, and azacytidine demethylation is a drug effect, not normal development.
How this study connects
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