DNMT3A and DNMT3B targeted mutations in hESC - interplay between non-coding RNAs and DNA methylation during human hematopoiesis
|Title||DNMT3A and DNMT3B targeted mutations in hESC - interplay between non-coding RNAs and DNA methylation during human hematopoiesis|
|Sponsor||European Union's Seventh Framework Programme (FP7)|
Associated cell lines
DNA methylation is an epigenetic modification that is important for numerous processes including regulation of gene expression and maintaining genomic integrity. DNMT3A and DNMT3B are two important nuclear DNA methyltransferases that catalyze the methylation of unmethylated and hemimethylated substrates. Mutations in DNMT3A are frequently found in blood disorders such as acute myeloid leukemia (AML) and strongly correlate with poor outcome of the disease. Mutations of DNMT3B in turn, have been associated with Immunodeficiency, Centromere instability and Facial anomalies (ICF) syndrome. There is also increasing evidence that DNA methyltransferases can be regulated by non-coding RNAs, and that those in turn those can change the DNA methylation pattern and influence the expression of other genes. Here we propose to inactivate DNMT3A and DNMT3B in human embryonic stem cells (hESCs) using the CRISPR/Cas9 system to further investigate their interplay with noncoding RNAs (nc-RNAs). Such cells will serve as an excellent tool for examining how the disruption of DNMT3A and DNMT3B affect the ability of stem cell to differentiate into all germ layers, and what their role in malignant hematopoiesis is, in particular with respect to how and which nc-RNAs are involved in these processes.