Identification of genes regulated by loss of RRM1 in Ewing sarcoma cells.

Ribonucleotide reductase (RNR), which is composed of RRM1 and RRM2 subunits, is the rate limiting enzyme in the synthesis of deoxyribonucleotides and required for DNA replication and DNA damage repair. We used a conditional knockout (CRISPR/Cas9) and rescue approach to target RRM1 in Ewing sarcoma cells and identify downstream pathways impacted by the loss of RNR activity. RNA-seq analysis was performed at times 0 and 48 hours after removal of doxycycline from cell culture media, which results in loss of the RRM1 protein. We performed RNA-seq in Ewing sarcoma cell lines with conditional knockout of RRM1.