CRISPR dCAS9 Based Epigenetic Editing in vitro
CRISPR dCAS9 Based Epigenetic Editing in vitro
Epigenetic changes occur during the entire life course to adapt to environmental changes. Lifestyle choices such as eating habits, physical activity or weight gain impact our epigenome and lead to alterations. A common epigenetic modification is the methylation of cytosine-guanine-rich DNA sequences, the so-called CpG islands, which has a negative effect on gene expression of the associated gene loci.
Based on our previous findings for obesity specific methylation pattern in human subcutaneous- and omental visceral adipose tissue, this project aims to identify possible epigenetic alterations that define a "point of no return" in obesity and comorbidities development. Therefore, we perform target gene specific methylation editing in adipocytes to elucidate functional consequences on adipocyte differentiation. Thus, we are testing the reversibility of methylation remodeling in human adipocytes by means of epigenetic editing. To do so, we are using CpG site specific methylation deposition or removal in pre-adipocytes using the CRISPR dCAS9 SunTaq model which binds several copies of the epigenetic modulators DNA methyltransferase 3A (DNMT3A) or Tet methylcytosine dioxygenase 1 (TET1).