OTAR2080_Cancer_NK_evasion_CRISPR

We are trying to identify genes that enable cancer cells to evade killing by Natural Killer cells or genes that make the cancer cells die faster. A panel of ovarian and lung cancer cell lines that stably express Cas9 are transduced with a genome wide CRISPR library generated from the Vienna BioCentre. After allowing essential genes to drop out for 14 days the cancer cells are co-cultured with Natural Killer cells that have been expanded from peripheral blood for a duration based on the basal sensitivity of the cells to killing. A parallel replicate is kept without Natural killer cells and the DNA is extracted and samples prepared for sequencing to identify the gRNA present. Various analysis is performed on the gRNA counts to identify those that are enriched or depleted.