Brunello genome-wide CRISPR screen data in WT and CDK8-knockout HAP1-Cas9 treated with ATRi (AZD6738) or DMSO
WT or CDK8 KO HAP1-Cas9-expressing cells were transduced at a multiplicity of infection of 0.3 and 500-fold library coverage of the Brunello human genome wide sgRNA library. 24 h post transduction, cells were selected with 1 µg/mL puromycin for 6 days to enrich for sgRNA expressing populations. Following selection, cells were split into two treatment arms; DMSO treated or treated with ATRi (AZD6738) at an IC20 for 10 population doublings. Cell pellets were collected at day 0 of treatment and at day 10 of treatment, genomic DNA was isolated using the QIAamp Blood Maxi Kit (Qiagen), and genome-integrated sgRNA sequences were amplified by polymerase chain reaction (PCR) using the Q5 Mastermix (New England Biolabs Ultra II) and i7 multiplexing barcoded primers. Products were sequenced on Illumina NovaSeq 6000 systems. Brunello sgRNA sequences were verified for off-target effects using Exorcise. Guide enrichment analysis was performed using DrugZ to compare DMSO-treated to ATRi-treated samples.