scJAK1screen

Immune checkpoint blockade (ICB) has emerged as one of the most promising therapeutic options for cancer treatment. ICB therapies inhibit pathways involved in cancer immune evasion, such as PD-L1-PD-1 and CTLA-4. However, some patients exhibit resistance (primary or acquired) which has been associated with inactivation of IFN? pathway components in tumour cells eg mutations in JAK1, a protein kinase downstream of the IFN? receptor. In this study we have screened JAK1 to identify the effects of variants of uncertain significance (VUS), using a cytosine base editor in HT29 cells. We have analysed edited cells at an early timepoint after treatment with IFN?, using both scRNAseq (10x) and scGenotyping (Mission Bio) to link together the identity of the edits with a transcriptomic readout.