Low-Cell-Number Epigenome Profiling Aids the Study of Lens Aging and Hematopoiesis

•RP-ChIP-seq enables high-fidelity epigenetic profiling in 500 cells•FARP-ChIP-seq is generally applicable•Age-associated epigenome changes in mouse lens are revealed by RP-ChIP-seq•Lack of H3K4me3/H3K27me3 bivalency on hematopoietic differentiation genes in HSCs

SummaryUnderstanding how chromatin modification regulates development and disease can be limited by available material. Despite recent progress, balancing high-quality and reliable mapping using chromatin-immunoprecipitation-based deep sequencing (ChIP-seq) remains a challenge. We report two techniques, recovery via protection (RP)-ChIP-seq and favored amplification RP-ChIP-seq (FARP-ChIP-seq), that provide reproducible mapping in as few as 500 cells. RP-ChIP-seq allows detection of age-associated epigenetic changes in a single mouse lens, whereas FARP-ChIP-seq accurately maps histone H3 lysine 4 trimethylation (H3K4me3) and H3K27me3 in long-term hematopoietic stem cells (LT-HSCs), short-term HSCs (ST-HSCs), and multi-potent progenitors (MPPs) from one mouse. These datasets not only highlight genes that may be involved in lens aging but also indicate a lack of H3K4me3/H3K27me3 bivalency on hematopoietic genes in HSCs.

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