Next Generation Sequencing Comparison of Wild Type and Whsc1-/- BM sorted proB Cell Transcriptomes. Mus musculus

Whsc1 gene codes for a SET domain-containing H3K36 dimethylase, whose activity has been suggested, in ex vivo cell culture experiments, to control many aspects of DNA and RNA processing (replication, repair, transcription, etc). Its precise function in vivo is still unclear. Here, we use RNA-seq transcriptome analysis to study the changes in gene expression in the absence of Whsc1 in sorted Bone Marrow proB cells. Our results show that, in the experimental system used, loss of Whsc1 caused massive changes in genes affecting many fundamental cellular processes and also severly interefering with B cell fate specification and commitment Overall design: Whsc1-KO mice are embryonic lethal. We therefore took hematopoietic cells from fetal liver of WT and Whsc1-KO embryo littermates and injected them in to lethally irradiated RAG1-KO recipients and allowed the generation of a full Whsc1-KO hematopoietic system. Flow cytometry analyses showed the existence of serious differentiation alterations in Whsc1-KO B cells. Then, we sorted B220+CD19+ckit+CD25-PI- proB cells form the BM of mice injected with either WT or Whsc1-/- cells andf performed RNAseq analyses of 2 independent WT and 2 independent Whsc1-KO biological replicates and we used these data to identify differentially expressed genes and pathways regulated by Whsc1 in pro-B cells.