Experimental Evolution of Cryptococcus neoformans in Mouse Macrophages

Interactions between fungal pathogens and mammalian immune cells can drive adaptation in fungal populations with important implications for disease progression and outcomes. To understand the ways these interactions can drive such adaptive evolution, we serially passaged the Cryptococcus neoformans strain Ftc555-1 through the mouse macrophage cell line, J774A.1. Fungal cells were passaged a total of 12 times through the macrophage cell line and were subsequently assessed for a change in their ability to replicate inside of macrophages. Populations and individual clonal cultures were identified with increased replication in macrophages and the genetic change underlying the adaptive phenotype of one of these populations was identified and tracked over the course of the evolution experiment by whole genome sequencing. High quality genomic DNA was extracted from lyophilized cell pellets using CTAB extraction protocols.