IFN-? induced transcriptional profile of A549 cells in absence of key transcriptional regulators IRF1 and NF2

Type-II interferon or IFN-gamma is a critical cytokine in innate immunity that control multitude of intracellular infections. IFN-gamma binds to interferon gamma receptors to phosphorylate and activate STAT1 mediated transcription of interferon stimulated genes (ISGs). We executed a genome-wide CRISPR-Cas9 screen against ~20,000 human genes to identify IRF-1 and NF2 as significant regulators of Toxoplasma infection in A549 lung epithelial cells. IRF-1 is a key transcription factor immediately downstream of STAT1 induced transcription that amplifies ISG expression in different cell types. NF2 or MERLIN is a tumor suppressor that is known to regulate expression of genes involved with cellular growth and metabolism. Transcriptomic signature of A549 cells lacking IRF-1 and NF2 were measured upon activation with IFN-gamma to find set of genes that may be regulating Toxoplasma infection. A total of 1,046,709,676 reads from 3 independent biological replicates were mapped to human hg38 reference genome. Expression of a ISGs that failed to induced in cells lacking IRF1 were found to be dysregulated in absence of NF2 as well. Overall design: Wild type, IRF1 KO and NF2 KO A549 cells were left untreated or treated with IFN-gamma treatment (100 U/ml) for 24 h. Total RNA was extracted from 3 independent biological replicates and submitted for next generation sequencing to analyze the role of IRF-1 and NF2 in IFN-gamma induced transcriptional profile of A549 cells.