Enhanced activities of OCT4 and SOX2 facilitates epigenetic reprogramming by modulating cell cycle [OSK/OvSvK CUT&TAG]
Fusing VP16  transcription activation domain with OCT4 and SOX2 (OvSvK) promoted the generation of induced pluripotency stem cells (iPSCs) by remodeling cell cycle and epigenetic modification. Multiomic analyses revealed that the other 80% upregulated genes experienced increase in H3K4me3 and decrease in H3K27me3 and were enriched with genes which undergo more intensive DNA demethylation during cell proliferation. Consistent results were obtained when siCcnd1, siCdkn2a and Ccne1 were screened out and used to mimic the cell cycle remodeling. In summary, these studies set up a correlation between epigenetic modification during cell cycle remodeling and cell fate conversion. Overall design: For bulk RNA-seq, cells were infected with O, S, K, OS, OK and SK respectively and collected at  post-infected Day0. For single-cell RNA-Seq, reprogramming cells were collected at post-infected Day0, Day1, Day2, Day3 and Day4 using Fluidigm C1. For ATAC-seq, cells were infected with hCdt1 and hGeminin and also OSK or OvSvK. Cells at different cell cycle stage were isolated by fluorescence-activated cell sorting (FACS). For CUT&Tag, cells infected with OSK or OvSvK and collected at post-infected Day3 and Day2 respectively.