Strains and Strategies for Large-Scale Gene Deletion Studies of the Diploid Human Fungal Pathogen Candida albicans

Noble, Suzanne M.; Johnson, Alexander D.

doi:10.1128/ec.4.2.298-309.2005

Cited by 592Fungal and yeast genetics researchAntifungal resistance and susceptibilityFungal Infections and Studies

Strains and Strategies for Large-Scale Gene Deletion Studies of the Diploid Human Fungal Pathogen Candida albicans

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Noble, Suzanne M., Johnson, Alexander D.·2005·Eukaryotic Cell

DOI 10.1128/ec.4.2.298-309.2005PMC PMC549318

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Abstract

Candida albicans is the most common human fungal pathogen and causes significant morbidity and mortality worldwide. Nevertheless, the basic principles of C. albicans pathogenesis remain poorly understood. Of central importance to the study of this organism is the ability to generate homozygous knockout mutants and to analyze them in a mammalian model of pathogenesis. C. albicans is diploid, and current strategies for gene deletion typically involve repeated use of the URA3 selectable marker. These procedures are often time-consuming and inefficient. Moreover, URA3 expression levels—which are susceptible to chromosome position effects—can themselves affect virulence, thereby complicating analysis of strains constructed with URA3 as a selectable marker. Here, we describe a set of newly developed reference strains ( leu2Δ/leu2Δ , his1Δ/his1Δ ; arg4Δ/arg4Δ , his1Δ/his1Δ ; and arg4Δ/arg4Δ , leu2Δ/leu2Δ , his1Δ/his1Δ ) that exhibit wild-type or nearly wild-type virulence in a mouse model. We also describe new disruption marker cassettes and a fusion PCR protocol that permit rapid and highly efficient generation of homozygous knockout mutations in the new C. albicans strains. We demonstrate these procedures for two well-studied genes, TUP1 and EFG1 , as well as a novel gene, RBD1 . These tools should permit large-scale genetic analysis of this important human pathogen.

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The auxotrophic markers HIS1 and ARG4 do not affect azole susceptibility of C. albicans in nutrient-rich media such as YPD.
“These auxotrophic markers do not affect azole susceptibility of C. albicans in nutrient-rich media such as an YPD medium, as used throughout this study [20,22].”
From Disruption of the UPC2 Gene Enhances Fluconazole Antifungal Activity by Inhibiting HAC1 mRNA Splicing in Candida albicans (2026) · cites this paper as [22]

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Method
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Authors: Noble, S. M., & Johnson, A. D.
Affiliations: University of California, San Francisco (US)
ORCID: Noble, Suzanne M.
Johnson, Alexander D.
Venue: Eukaryotic Cell
Publisher: American Society for Microbiology
Published: 2005-02
Volume: 4
Issue: 2
Pages: 298-309
ISSN: 1535-9778 (ISSN-L)
1535-9786
DOI: 10.1128/ec.4.2.298-309.2005
PMC: PMC549318
OpenAlex: W2128207311
Semantic Scholar: 858fb504935ca0d20038974fccfd87dc6493a317
Cited by: 592
References: 57
Citations by year: 2026: 22 · 2025: 21 · 2024: 39 · 2023: 16 · 2022: 35 · 2021: 27 · 2020: 28 · 2019: 35 · 2018: 33 · 2017: 36 · 2016: 35 · 2015: 46 · 2014: 36 · 2013: 32 · 2012: 23
FWCI: 6.53
Type: journal article
Language: en
Open access: diamond
doi.org
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Eukaryotic Cell (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health
National Institute of Allergy and Infectious Diseases (R01AI049187, R01 AI 49187)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
Health Sciences › Medicine › Epidemiology › Fungal Infections and Studies
Keywords: Candida albicans, gene deletion, fusion PCR, reference strains, virulence, Tup1, EFG1
MeSH terms: Animals, Candida albicans, Fungal Proteins, Genes, Fungal, Genetic Complementation Test, Genetic Markers, Humans, Gene Expression Regulation, Fungal, Survival Rate, Polymerase Chain Reaction, Gene Deletion, Mice

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Strains and Strategies for Large-Scale Gene Deletion Studies of the Diploid Human Fungal Pathogen Candida albicans

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