Monitoring Phenotypic Switching in Candida albicans and the Use of Next-Gen Fluorescence Reporters

Curr Protoc Microbiol. 2019 Jun;53(1):e76. doi: 10.1002/cpmc.76. Epub 2019 Feb 12.

Abstract

Candida albicans is an opportunistic human fungal pathogen that is able to cause both mucosal and systemic infections. It is also a frequent human commensal, where it is typically found inhabiting multiple niches including the gastrointestinal tract. One of the most remarkable features of C. albicans biology is its ability to undergo heritable and reversible switching between different phenotypic states, a phenomenon known as phenotypic switching. This is best exemplified by the white-opaque switch, in which cells undergo epigenetic transitions between two alternative cellular states. Here, we describe assays to quantify the frequency of switching between states, as well as methods to help identify cells in different phenotypic states. We also describe the use of environmental cues that can induce switching into either the white or opaque state. Finally, we introduce the use of mNeonGreen and mScarlet fluorescent proteins that have been optimized for use in C. albicans and which outperform commonly used fluorescent proteins for both fluorescence microscopy and flow cytometry. © 2019 by John Wiley & Sons, Inc.

Keywords: Candida albicans; cell identity; fluorescent reporters; gene expression; heritable states; phenotypic switching.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Candida albicans / chemistry
  • Candida albicans / genetics*
  • Candida albicans / growth & development
  • Candida albicans / metabolism
  • Colony Count, Microbial / methods*
  • Culture Media / chemistry
  • Culture Media / metabolism
  • Flow Cytometry / methods*
  • Genes, Reporter
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Microscopy, Fluorescence / methods*
  • Phenotype
  • Polymerase Chain Reaction / methods*
  • Transformation, Genetic

Substances

  • Culture Media
  • Luminescent Proteins