CRAC of yeast RNA polymerase II in Topoisomerase 1 and 2 depleted cells

We obtained RNA polymerase II profiles across the genome of S.cerevisiae cells harbouring Top1-AID and Top2-AID tagged proteins. Cells were treated with anauxin analog (IAA) to induce degradation of Top1-AID and Top2-AID. This allowed us to compare RNAPII in absence of Top1 and Top2 activity to the control condition (absence of IAA treatment). The nascent RNA was crosslinked to Rpb1 (RNAPII) and sequenced after purification of the target protein under stringent and denaturing conditions. This reveals the position of the target protein at the nucleotide resolution level.