A role of p53 in mediating epigenetic stress (DRIPc-seq)

5-Azacytidine (Aza) and 5-Aza-2'-deoxycytidine (Dac) are the only epigenome modifying drugs in widespread clinical use, and are some of the primary therapeutics for Acute Myeloid Leukaemia treatment. The two are frequently used interchangeably, surprisingly as their selectivity of de-methylation is unique from the other, with to date no identified predictors of response or known biomarkers to indicate drug preference. Using these drugs to induce de-methylating conditions, we combine data from DRIPc-Seq, Immunostaining, RNA-Seq and Mass spectrometry with epigenetic analysis to uncover unique cellular responses following Aza and Dac treatment. HEK293 (WT) and p53 KO HEK293 (p53KO) cells were treated with 1.5 µM of Azacitidine or Decitabine or solvent for 48 hours and DRIPc-seq was performed in two replicates to assess the change to the formation of R-loops. Two additional HEK293 (WT) control samples were pre-treated with RNaseH, which specifically removes R-loops.