Chrystal Thomas

Candida albicans comprises a significant portion of the human microbiome, and when invasive, can cause high rates of morbidity and mortality. However, little is known about the interaction between C. albicans and the human immune system, and in particular, the transcriptional networks underlying the outcome of these interactions. In order to understand the factors that determine whether the immune system will successfully suppress C. albicans invasion, we worked on developing a novel set of microscopy and RNA-seq tools. Central to this effort is the ability to fix (or, freeze in time) the interactions between C. albicans and host immune cells in vitro. We therefore explored different fixation and RNA isolation methods. The two main criteria for our fixative were (1) to preserve the fluorescence signature of host and pathogen cells for flow cytometry and sorting and (2) to preserve the RNA integrity of cells for subsequent sequencing of sorted subpopulations via RNA-seq. We discovered that RNAlater, a commercially available RNA preservation solution, met both of our criteria and is a promising candidate for further study. This work will help facilitate the development of an imaging and RNA-seq analysis pipeline to dissect the mechanistic and transcriptional basis of infection outcome. Ultimately, these studies will aid in the identification of targets for therapeutic intervention.

PROJECT:Novel methodology to identify host and pathogen determinants of Candida albicans infection

Mentor: Dawn Thompson, Cell Circuits Program