Whole genome sequencing of RPE1-hTERT-DAAO^H2B P53 KO and RPE1-hTERT-DAAO^TOM20 P53KO cells
RPE1-hTERT p53KO cells stably expressing D-amino acid oxidase (DAAO) located at the histones (RPE1-hTERT-DAAO^H2B) or DAAO located to the outer mitochondrial membrane (RPE1-hTERT-DAAO^TOM20) were generated. This allows for localized H2O2 production in the nucleus or outer mitochondrial membrane of mitochondria upon addition of D-Alanine. L-Alanine was used as a control compound. These lines were treated for 2h 20 mM L-Ala or D-Ala per day for 4 consecutive days, cultured for an additional 3 days, then monoclonally sorted by FACS to allow for monoclonal outgrowth. Genomic DNA of of clones was harvested by use of the DNeasy blood and tissue kit (Qiagen). WGS libraries were generated using standard protocol (Illumina), which were sequenced to 15x coverage (2 x 150 bp) on an illumina Novaseq 6000 machine.