Large‐scale essential gene identification in Candida albicans and applications to antifungal drug discovery

  title={Large‐scale essential gene identification in Candida albicans and applications to antifungal drug discovery},
  author={Terry Roemer and Bo Jiang and John Davison and Troy Ketela and Karynn Veillette and Anouk Breton and Fatoumata Tandia and Annie Linteau and Susan Sillaots and Catarina Marta and Nick Martel and Steeve Véronneau and S{\'e}bastien Lemieux and Sarah J Kauffman and Jeffrey M. Becker and Reginald Storms and Charles Boone and Howard Bussey},
  journal={Molecular Microbiology},
Candida albicans is the primary fungal pathogen of humans. Despite the need for novel drugs to combat fungal infections [Sobel, J.D. (2000) Clin Infectious Dis 30: 652], antifungal drug discovery is currently limited by both the availability of suitable drug targets and assays to screen corresponding targets. A functional genomics approach based on the diploid C. albicans genome sequence, termed GRACETM (gene replacement and conditional expression), was used to assess gene essentiality through… 

Gene Annotation and Drug Target Discovery in Candida albicans with a Tagged Transposon Mutant Collection

The value of haploinsufficiency screens directly in this pathogen for gene annotation and drug target identification is highlighted and constructed and screened a library of 3633 tagged heterozygous transposon disruption mutants, using them in a competitive growth assay to examine nutrient- and drug-dependent haplo insufficiency.

Conserved Fungal Genes as Potential Targets for Broad-Spectrum Antifungal Drug Discovery

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Functional genomic approaches to fungal pathogenesis, drug target validation, and antifungal drug discovery.

The availability of a complete barcoded set of deletion mutants for the major fungal pathogens would permit identification of the “universal fungal essential gene set,” allow a broad application of chemical genomics in these organisms, and enable a systematic analysis of phenotypes associated with pathogenicity.

Genome-Wide Fitness Test and Mechanism-of-Action Studies of Inhibitory Compounds in Candida albicans

Candida albicans is a prevalent fungal pathogen amongst the immunocompromised population, causing both superficial and life-threatening infections. Since C. albicans is diploid, classical

Advances in fungal chemical genomics for the discovery of new antifungal agents

Current yeast chemical genomic assays are examined and it is highlighted how such resources provide powerful tools that can be utilized to bolster the antifungal pipeline.

Essential Gene Identification and Drug Target Prioritization in Aspergillus fumigatus

It is demonstrated the pNiiA-CPR strategy is suitable for in vivo phenotypic analyses, as a number of conditional mutants, including an ERG11 double mutant, failed to establish a terminal infection in an immunocompromised mouse model of systemic aspergillosis.

Essential Gene Discovery in the Basidiomycete Cryptococcus neoformans for Antifungal Drug Target Prioritization

The identification of C. neoformans genes that are essential for viability as identified through forward and reverse genetic approaches, using an engineered diploid strain and genetic segregation after meiosis, represent an excellent starting point for the future development of new antifungals by pharmaceutical companies.

DeORFanizing Candida albicans Genes using Coexpression

The utility of this coexpression network is demonstrated in both retrospective and prospective testing and CalCEN is used to predict a role for C4_06590W/CCJ1 in cell cycle and to allow for a better characterization of underannotated genes in pathogenic yeasts.

A Versatile Overexpression Strategy in the Pathogenic Yeast Candida albicans: Identification of Regulators of Morphogenesis and Fitness

The development of a first generation C.Albicans ORFeome, the improvement of overexpression systems and the construction of two new libraries of C. albicans strains overexpressing genes for components of signaling networks, in particular protein kinases, protein phosphatases and transcription factors are reported.

Comparative genomics allowed the identification of drug targets against human fungal pathogens

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A genome‐wide strategy for the identification of essential genes in Staphylococcus aureus

A cell‐based, drug‐screening assay is described, wherein expression of an antisense RNA confers specific sensitivity to compounds targeting that gene product, thereby greatly facilitating the search for new antibiotics.

Recent developments in molecular genetics of Candida albicans.

This review outlines recent advances in the development of molecular tools for functional analysis in C. albicans and summarizes current knowledge about the genomic and genetic variability of this important human fungal pathogen.

Identification of the FKS1 gene of Candida albicans as the essential target of 1,3-beta-D-glucan synthase inhibitors

It is concluded that C. albicans strains CAI4R1, NR2, and NR4 are heterozygous for a dominant or semidominant pneumocandin resistance mutation at CaFKS1, and CaFks1p is a target of the echinocandins.

Regulation of both gene expression and protein stability provides genetically assisted target evaluation (GATE) for microbial target validation.

GATE concomitantly manipulates both synthesis and stability of the targeted protein using copper ions as an effector and allows rapid quantitation of the lethal consequences of inactivation of targeted gene products in Saccharomyces cerevisiae.

Genomic Profiling of the Response of Candida albicans to Itraconazole Treatment Using a DNA Microarray

This report is the first to describe the application of DNA chip technology to study the response of a major human fungal pathogen to drug treatment and might provide—in combination with phenotypic data—first hints of their potential function.

Signature‐tagged and directed mutagenesis identify PABA synthetase as essential for Aspergillus fumigatus pathogenicity

The successful application of signature‐tagged mutagenesis to Aspergillus fumigatus demonstrates that in vivo genetic analysis of eukaryotic pathogens is feasible and could result in the identification of potential targets, such as para‐aminobenzoic acid synthetase, for novel antifungal therapies.

Single-Copy IMH3 Allele Is Sufficient To Confer Resistance to Mycophenolic Acid in Candida albicans and To Mediate Transformation of Clinical Candida Species

It is demonstrated that the wild-typeIMH3 gene, when expressed in high copy number, will confer resistance to mycophenolic acid and cloned and sequenced the IMH3r gene from C. albicans strain 1006 has three amino acid changes, two of which are nonconservative, and it is shown that at least two of the three mutations are required to conferred resistance to MPA.

A hyphal-specific chitin synthase gene (CHS2) is not essential for growth, dimorphism, or virulence of Candida albicans.

  • N. GowP. Robbins O. Kinsman
  • Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 1994
In the dimorphic fungus Candida albicans, the CHS2 gene encodes a chitin synthase that is expressed preferentially in the hyphal form and the "ura-blaster" protocol described for Saccharomyces, which involves the sequential disruption of multiple alleles by integrative transformation with URA3 as a single selectable marker, was achieved.

A Single-Transformation Gene Function Test in DiploidCandida albicans

A genetic construct (UAU1) for which two copies may be selected, insertion of UAU1 into one genomic site, allows selection for segregants with two copies of the insertion to assess rapidly the essentiality of C. albicans genes.

Tetracycline-Regulatable System To Tightly Control Gene Expression in the Pathogenic Fungus Candida albicans

A convenient system to control gene expression in C. albicans by the tetracycline-regulatable (TR) promoters is developed, able to function in both in vitro and in vivo settings, and is useful for investigating gene functions.