Npac regulates pre-mRNA splicing in mouse embryonic stem cells

As a reader of tri-methylated lysine 36 on histone H3 (H3K36me3), Npac has been shown a significant role in gene transcription elongation. However, its potential implication in RNA splicing remains unknown. Here, we characterized the phenotypes of Npac knockout in mES cells. We discovered that loss of Npac disrupts pluripotency and identity in mESCs. We also found that Npac is associated with many cellular activities including cell proliferation, differentiation, transcription regulation. Notably, we uncovered that Npac is associated with RNA splicing machinery. Furthermore, we found that Npac regulates alternative splicing through its interaction with the splicing factors including Srsf1. Our research thus highlights the important role of Npac in maintaining ESC identity through regulation of pre-mRNA splicing. Overall design: To elucidate the cooperative role of Npac in regulating mouse embryonic stem cells (mESCs), we generated E14 cell lines in which specific target genes were knocked out using CRISPR-Cas9 technology. Following this, we performed a comparative gene expression profiling analysis by employing RNA-sequencing. This process enables a comprehensive overview of the RNA-seq data analysis, comparing the gene expression patterns between the wild-type (WT) E14 cells and their knockout (KO) derivatives.