Detection of DNA double-strand breaks in T-47D cells using DSBCapture-seq

Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3B, A3B) is a key molecular driver inducing mutations in multiple human cancer. A3B belongs to the APOBEC3 enzyme family, which consists seven closely related DNA deaminases that catalyse cytosine-to-uracil (C>U) editing of single-stranded DNA (ssDNA). Here, we used DSBCapture-seq to study the formation of DNA double-strand breaks (DSB) by estrogen receptor activation upon 17-estradiol (E2) induction. The effect of A3B-depletion on DSB formation was investigated using RNA interference. T-47D cells were divided into three groups and treated with siNT+DMSO, siNT+E2 and siA3B+E2, where siNT and siA3B and denotes non-targeting and A3B-targeting siRNA respectively. Experiments were carried out with two biological repeats.