|Sponsor||European Research Council (ERC)|
|Institution||Wolf Reik Lab|
Associated cell lines
Dppa2 and 4 are small chromatin bound proteins exclusively expressed during pluripotency in mouse. We found that they are needed in mouse ESCs to target bivalency (H3K4me3 and H3K27me3) to a subset of bivalent promoters. In the absence of Dppa2 and 4 bivalency and chromatin accessibility is lost, and DNA methylation gained, and developmental genes fail to be expressed during differentiation (Eckersley-Maslin et al Nature Structural and Molecular Biology 2020). DPPA2 and 4 are conserved in human and in this project we propose to characterise the expression and their role in human pluripotency and differentiation. DPPA2 and 4 will be deleted in hESCs by Crispr/Cas9 targeting, and transcriptional and epigenetic properties of the cells will be studied during the pluripotent state and during in vitro differentiation. We have discovered that neuroectoderm enhancers (but not mesodermal or endodermal ones) are primed by chromatin accessibility and hypomethylation in mouse embryonic stem cells (Argelaguet et al 2019 Nature). In this project we aim to determine if such priming also exists in human embryonic stem cells, and if so, what its function might be in neuroectoderm differentiation in vitro. We will use enhancer annotations from various in vitro differentiation and from in vivo tissue datasets, and map their epigenetic properties in hESCs. We will subsequently manipulate such elements in hESCs by Crispr/Cas9 targeting, and determine the impact of altering their function on neuroectoderm commitment and differentiation in vitro.