Fluorescent organelle markers in Cryptococcus neoformans : a versatile toolkit for live-cell subcellular localization.

Understanding how intracellular organelles are organized and dynamically remodeled is essential for elucidating fungal growth, differentiation, and pathogenicity. The human fungal pathogen Cryptococcus neoformans exhibits remarkable morphological diversity, yet the contribution of intracellular orga...

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Publicado en:Genetics Vol. 233; no. 3; pp. 1 - 16
Autores principales: Choi, Yeseul, Bahn, Yong-Sun, Heitman, Joseph
Formato: pictorial research tables/charts Journal Article
Publicado: Oxford University Press / USA Jul2026
Acceso en línea:Ver este registro en EBSCOhost
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      pub: Oxford University Press / USA
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        10.1093/genetics/iyag110
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        atl: Fluorescent organelle markers in Cryptococcus neoformans : a versatile toolkit for live-cell subcellular localization.
      aug:
        au:
          Choi, Yeseul
          Bahn, Yong-Sun
          Heitman, Joseph
        affil: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Duke University, Durham, NC 27710, United States
      sug:
        subj:
          Cryptococcus Analysis
          Cell Physiology
          Fluorescent Dyes
          Organelles
          Biological Markers
          Luminescent Measurements
          Human
          Funding Source
          Male
          Female
          Adult
          Middle Age
          Endoplasmic Reticulum
          Cytoplasm
          Cell Differentiation
          Plasmids
          Cell Nucleus
          Mitochondria
          Autophagy
          Cell Membrane
          Cell Division
          Reverse Transcriptase Polymerase Chain Reaction
          Clustered Regularly Interspaced Short Palindromic Repeats
          CRISPR-Associated Proteins
          Electroporation
          Post Hoc Analysis
          Microscopy
          Staining and Labeling
          One-Way Analysis of Variance
          In Vitro Studies
          Descriptive Statistics
          Adult: 19-44 years
          Middle Aged: 45-64 years
          Male
          Female
      ab: Understanding how intracellular organelles are organized and dynamically remodeled is essential for elucidating fungal growth, differentiation, and pathogenicity. The human fungal pathogen Cryptococcus neoformans exhibits remarkable morphological diversity, yet the contribution of intracellular organelles to adaptation across environmental and host-relevant conditions have not been systematically examined. Here, we establish a comprehensive toolkit of fluorescent organelle marker plasmids and strains expressing mCherry or GFP fusions, enabling high-resolution live-cell imaging of 10 major subcellular compartments: the nucleolus, endoplasmic reticulum (ER), Golgi apparatus, mitochondria, peroxisome, endosome, autophagosome, vacuole, the plasma membrane, and processing bodies (P-bodies). With this platform, we analyzed organelle organization under host-relevant conditions, including elevated temperature, 5% CO2, and capsule- and melanin-inducing media, as well as throughout sexual development from zygote formation to basidiospore production. We further show that the toolkit supports colocalization analyses via diploid formation or dual-labeling strategies. Collectively, our results reveal condition- and stage-specific remodeling of organelle architecture during vegetative growth and mating. This imaging platform provides a robust framework for investigating how subcellular organization supports fungal adaptation, development, and virulence, and for inferring the functions of uncharacterized genes from their spatial localization.
      pubtype: Academic Journal
      doctype:
        pictorial
        research
        tables/charts
        Journal Article
      ougenre: Article
    language: English
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