Haploid Screening for the Identification of Host Factors in Virus Infection

Elucidating which host factors are exploited by viruses to infect target cells is key to our understanding of how these pathogens cause disease and how it might be counteracted by future therapies. Pooled gene-trap mutagenesis of haploid human HAP1 cells has proven to be a formidable tool for revealing genes involved in the infection process for a suite of human pathogenic viruses. This method has led to the identification of a number of virus receptors and unconventional entry mechanisms into human cells. In the case of Ebola virus, for example, the discovery of the lysosomal protein NPC1 as an intracellular receptor sparked the development of tailored strategies to interfere with viral infection. The "single tube" pooled screening technique presented here does not require any automation or robotics and is potentially applicable to any virus able to infect HAP1 cells.