Comparision of Differential methylation between Control and mutant Dnmt3a

Dnmt3a is the most recurrently mutated gene in clonal hematopoiesis (CH), and it is a critical regulator of hematopoietic stem cells (HSCs). Conditional deletion of Dnmt3a in mouse HSC results in enhanced self-renewal but impaired differentiation. Dnmt3a encodes for a de novo DNA methyltransferase enzyme but both mouse and human cells with loss of Dnmt3a show minimal change in DNA methylation levels which do not correlate with gene expression differences. To understand if there are methylation differences between control and mutant (R878H) Dnmt3a, we are performing WGBS. To investigate the differential DNA methylation in hematopoietic cells expressing Dnmt3a mutant with reduced DNA methyltransferase activity, we established Cre-mediated knock in mouse model. Competitive HSC transplantation was performed. Cells were isolated 18-weeks post-transplant for analysis of DNA methylation patterns by whole genome bisulfite sequencing.