Cytokine Stimulation of PTPN2-deleted cancer cells

PTPN2 was deleted from a selection of murine and human cancer cells using CRISPR/Cas9. The loss-of-function phenotype was assessed in vitro with cytokine stimulation or vehicle control. In order to determine the effect of PTPN2-inhibition of interferon signalling, Cas9-expressing murine (B16 melanoma) and human (A375, A549, HT29, MelJuSo) cancer cells were infected with lentivirus containing guide RNA (sgRNA) against Ptpn2 or a non-targeting sequence. After Ptpn2 deletion was verified, the cell lines were treated with vehicle control (PBS with 0.1% BSA), interferon-beta (IFNb), interferon-gamma (IFNg), or tumor necrosis factor (TNFa), alone or in combination, and transcriptionally profiled by directional, paired-end RNAseq after a 48 hour stimulation.!Series_overall_design = Please note that 'Bro3' and 'Bro4' refer to non-targeting guides provided by the Broad Institute. Please note that 'Bro3' and 'Bro4' refer to non-targeting guides provided by the Broad Institute.