Nascent RNA sequencing (PRO-seq) after rapid degradation of AID-tagged ZNF143

These experiments identify the primary response genes of ZNF143 in HEK-293T cells and the transcriptional response that results from treating cells with 500μM auxin. Oryza sativa TIR1 was inserted into all copies of the AAVS1 locus in HEK-293T cells and a single clone was expanded. All subsequent genetic modifications were derived from this progenitor cell line. Genetic modifications include endogenously tagging ZNF143 with an auxin-inducible degron (AID) and insetion of Auxin Response Factor (OsARF16). Cells were treated with 500μM auxin for 90 minutes to degrade ZNF143. Progenitor cells were treated with 500μM Auxin for 3 hours as a control experiment.