Project description:Mar1 deletion and RNA enrichment in Cryptococcus neoformans: pilot data for a high-throughput sequencing course. The goal of this project was to generate pilot data in preparation for a summer course on high-throughput sequencing where participants prepared their own RNA-Seq libraries and analyzed the resulting data. This pilot experiment addressed two questions: 1. Does this experimental system (Cryptococcus neoformans H99 wildtype and mar1 deletion mutant grown in YPD and tissue culture media) provide a good dataset for course participants to analyze. 2. Which rRNA depletion method is best to use in the wetlab component of the course. This data was generated in preparation for the intensive summer course on high-throughput sequencing, funded by NIH grant 5R25EB023928-03 "A hands-on, integrative next-generation sequencing course: design, experiment, and analysis".
Project description:Cryptococcus neoformans lab strain H99 was spread on YPD plate supplemented with 4ug/ml benomyl. Randomly 60 adaptors were sequenced.
Project description:Cryptococcus neoformans lab strain H99 was spread on YPD plate supplemented with 2.5ug/ml chlorothalonil. Randomly 72 adaptors were sequenced.
Project description:Cryptococcus neoformans is the most common cause of fungal meningitis, with high mortality and morbidity. The reason for the frequent occurrence of Cryptococcus infection in the central nervous system (CNS) is poorly understood. In this study, we find that inositol plays an important role in the transversal of Cryptococcus across the blood-brain barrier (BBB) both in an in vitro human BBB model and in vivo animal models. The inositol stimulation of BBB crossing is dependent upon fungal inositol transporters. The upregulation of genes involved in the inositol catabolism pathway is evident in a microarray analysis. The expression of CPS1, a gene encoding the hyaluronic acid synthase in Cryptococcus, is also upregulated by the inositol treatment. The production of hyaluronic acid increased in cells treated with inositol, which leads to the enhanced binding ability of Cryptococcus cells to the human brain microvascular endothelial cells (HBMECs) constituting the BBB. Overall, our studies provide a mechanism for inositol-dependent Cryptococcus transversal of the BBB, supporting our hypothesis that host inositol utilization by the fungus contributes to Cryptococcus CNS infection.
Project description:Cryptococcus neoformans lab strain H99 was spread on YPD plate supplemented with 1.5ug/ml pyraclostrobin. Randomly 23 adaptors were chosen. These adaptors were sequenced.
Project description:Cryptococcus neoformans lab strain H99 was spread on YPD plates supplemented with 32ug/ml and 64ug/ml iprodione. Randomly 29 (T2590-T2618( and 30 (T2619-2648) adaptors from 32ug/ml and 64ug/ml drug plate, respectively, were sequenced.
