Human fungal pathogens cause more than 1.5 million deaths annually. In particular, Cryptococcus species are responsible for life threatening meningitis and kill nearly 200,000 people a year. There are only three classes of effective antifungal drugs available to treat Cryptococcus and developing new drugs is difficult because many potential antifungal drugs are also toxic to humans. Effective use of this small set of drugs and development of new or complementary drugs will be aided by a better understanding of the genetic basis of both virulence and drug resistance. My lab will utilize high-throughput genetic/genomic approaches to identify interesting drug resistance and susceptibility relevant pathways and follow up on those discoveries with targeted molecular biology analyses. We developed a transposon mutagenesis approach for Cryptococcus that uses massively parallel in vivo transposition coupled with targeted high-throughput sequencing (TN-seq) to determine how every gene affects growth in a given condition in a single experiment. This assay allows us to rapidly determine the genes required for resistance and susceptibility to an antifungal agent in Cryptococcus neoformans. More broadly, TN-seq will allow my research group to interrogate evolution of virulence across the Cryptococcus pathogenic species complex, including avirulent sister species.