Analysis of gene expression in filamentous cells of Candida albicans grown on agar plates

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Abstract
Candida albicans (C. albicans) is a commensal organism of the human gastrointestinal and genitourinary tracts. C. albicans is also a major human pathogen, causing disease ranging from cutaneous infections to lethal systemic disease. The ability of this fungus to switch between yeast and filamentous forms of growth has long been linked to its pathogenesis. Filamentation can be induced by a variety of distinct environmental cues and can occur in either liquid or solid media. While some evidence suggests that there are differences between filamentation in solid and liquid media, gene expression analysis of filamentation in C. albicans has focused strictly on cells grown in liquid media. We have developed a method for analyzing gene expression of filamentous cells grown on solid induction media at early stages of filamentation, establishing cell plating densities, ideal collection times, and collection techniques. We have also demonstrated the utility of the approach not only in qRT-PCR assays, but high-throughput RNAseq assays as well. These assays will allow for comparison studies of C. albicans filamentation initiation in solid and liquid media.

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