Genome-wide CRISPR knockout screen in Mycobacterium tuberculosis infected macrophages to identify regulators of TNF and iNOS expression

To elucidate the early regulators of inflammation during Mycobacterium tuberculosis infection, we performed a genome-wide CRISPR knockout screen in macrophages to identify genes that influence the induction of TNF and iNOS upon infection. A genome-wide knockout library in murine macrophages was infected with Mycobacterium tuberculosis and 24 hours post infection cells were FACS sorted based on TNF and iNOS expression. Samples were collected from the unsorted library, as well as two sorted populations: TNF positive and iNOS negative, and TNF positive and iNOS positive.