CRISPR screen for genes involved in nuclear factor kappa B signaling activation by flagellin

Escherichia coli represents the most common Gram-negative organism causing neonatal meningitis. How human brain microvascular endothelial cells (HBMEC) interacts with the meningitis-causing E. coli remains incompletely understood. We observed that the signaling through nuclear factor kappa B was upregulated in HBMEC infected with E. coli. We demonstrated that the bacterial flagellin was the major contributor to nuclear factor kappa B activation in HBMEC. We performed a fluorescence-activated cell sorting (FACS)-based genome-wide CRISPR screen using the nuclear factor kappa B GFP reporter cell and identified several known genes as well as new genes required for nuclear factor kappa B activation by E. coli.