Genetic manipulation of the pathogenic yeast Candida parapsilosis

Nosek, Jozef; Adamíková, L'ubica; Zemanová, Júlia; Tomáška, Ľubonír; Zufferey, Rachel; Mamoun, Choukri Ben

doi:10.1007/s00294-002-0326-7

Cited by 32 publications

(30 citation statements)

References 49 publications

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“…Molecular genetic studies of C. parapsilosis have been hindered by the aneuploidy of the organism (22) and the lack of a characterized sexual cycle. A transformation system based on the complementation of a galactokinase-deficient mutant of C. parapsilosis by the homologous gene (GAL1) has been described, but it lacks efficiency and cannot be used to inactivate target genes in prototrophic WT strains (23). Our studies demonstrate that the SAT1 flipper cassette is an efficient tool to generate homozygous KO mutants in C. parapsilosis.…”

Section: Discussionmentioning

confidence: 89%

Targeted gene deletion in Candida parapsilosis demonstrates the role of secreted lipase in virulence

Gácser¹,

Trofa²,

Schäfer³

et al. 2007

J. Clin. Invest.

135

136

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Candida parapsilosis is a major cause of human disease, yet little is known about the pathogen's virulence. We have developed an efficient gene deletion system for C. parapsilosis based on the repeated use of the dominant nourseothricin resistance marker (caSAT1) and its subsequent deletion by FLP-mediated, site-specific recombination. Using this technique, we deleted the lipase locus in the C. parapsilosis genome consisting of adjacent genes CpLIP1 and CpLIP2. Additionally we reconstructed the CpLIP2 gene, which restored lipase activity. Lipolytic activity was absent in the null mutants, whereas the WT, heterozygous, and reconstructed mutants showed similar lipase production. Biofilm formation was inhibited with lipase-negative mutants and their growth was significantly reduced in lipid-rich media. The knockout mutants were more efficiently ingested and killed by J774.16 and RAW 264.7 macrophage-like cells. Additionally, the lipase-negative mutants were significantly less virulent in infection models that involve inoculation of reconstituted human oral epithelium or murine intraperitoneal challenge. These studies represent what we believe to be the first targeted disruption of a gene in C. parapsilosis and show that C. parapsilosis-secreted lipase is involved in disease pathogenesis. This efficient system for targeted gene deletion holds great promise for rapidly enhancing our knowledge of the biology and virulence of this increasingly common invasive fungal pathogen.

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Section: Discussionmentioning

confidence: 89%

Targeted gene deletion in Candida parapsilosis demonstrates the role of secreted lipase in virulence

Gácser¹,

Trofa²,

Schäfer³

et al. 2007

J. Clin. Invest.

135

136

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show abstract

“…Furthermore, until 2007 an efficient method for targeted gene disruption to generate mutants for the identification of C. parapsilosis virulence factors did not exist. A transformation system based on the complementation of a galactokinase-deficient mutant of C. parapsilosis by the homologous gene (GAL1) was described in 2002, but it could not be used to target genes in prototrophic wild-type strains (191). We previously showed that the dominant selection marker MPA R can be used for C. parapsilosis transformation (94), and we created an improved system for efficient gene deletion based on the repeated use of the dominant nourseothricin (Nou) resistance marker (CaSAT1) and its subsequent deletion by site-specific recombination (97) as previously described for C. albicans (227).…”

Section: Molecular Manipulationsmentioning

confidence: 99%

Candida parapsilosis, an Emerging Fungal Pathogen

2008

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SUMMARY Candida parapsilosis is an emerging major human pathogen that has dramatically increased in significance and prevalence over the past 2 decades, such that C. parapsilosis is now one of the leading causes of invasive candidal disease. Individuals at the highest risk for severe infection include neonates and patients in intensive care units. C. parapsilosis infections are especially associated with hyperalimentation solutions, prosthetic devices, and indwelling catheters, as well as the nosocomial spread of disease through the hands of health care workers. Factors involved in disease pathogenesis include the secretion of hydrolytic enzymes, adhesion to prosthetics, and biofilm formation. New molecular genetic tools are providing additional and much-needed information regarding C. parapsilosis virulence. The emerging information will provide a deeper understanding of C. parapsilosis pathogenesis and facilitate the development of new therapeutic approaches for treating C. parapsilosis infections.

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“…Expression studies of CpLIP genes using other yeasts highly contributed to our knowledge on C. parapsilosis lipase secretion, but did not address their role in virulence. Although there were advances in C. parapsilosis genetic manipulation [12,54] , it was not until 2007 that an innovative method for targeted gene deletion became available, and LIP1 -LIP2 were one of the first genes to be deleted from the diploid genome of C. parapsilosis, using a dominant selection marker carrier flipper cassette [34]. The available null mutant strain (Cp ΔΔlip1-ΔΔlip2) provided us a novel tool for studying C. parapsilosis lipases and facilitated numerous subsequent virulence related investigations.…”

Section: Identification Of C Parapsilosis Secreted Lipasesmentioning

confidence: 99%

Candida parapsilosis Secreted Lipase as an Important Virulence Factor

Tóth

Vágvölgyi

et al. 2017

CPPS

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Abstract:The prevalence of Candida parapsilosis, an opportunistic human pathogenic fungal species, is increasing at an alarming rate in the hospital environment. Patients at risk for C. parapsilosis infection include those with immunosuppression, such as individuals with cancer, AIDS, and low birth weight premature neonates as well as patients that had undergone abdominal surgery. Neonatal candidiasis caused by C. parapsilosis has been widely reported across the globe. Various reports have shown that, compared to other Candida species, certain C. parapsilosis clinical isolates were less susceptible to antifungals such as amphotericin B, fluconazole, and caspofungin. In addition, some studies have even reported multi-echinocandin or multi-azole resistant strains of C. parapsilosis. C. parapsilosis has several virulence factors that contribute to its capacity for host invasion and among these factors extracellular lipases have a major role in pathogenesis. In this review we have collected all the recent relevant studies that confirm the involvement of secreted lipases in C. parapsilosis pathogenesis, using both in vitro and in vivo models of infection. Of particular note, an available lipase deficient C. parapsilosis strain has been utilized to demonstrate that the lack of secreted lipases decreased virulence, reduced tissue damage, and was less able to survive within phagocytes or mice compared to the wild type. Since fungal secreted lipases have different characteristics than lipolytic enzymes present in humans, C. parapsilosis extracellular lipases may be potential targets for the development of novel antifungal drugs.

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Genetic manipulation of the pathogenic yeast Candida parapsilosis

Cited by 32 publications

References 49 publications

Targeted gene deletion in Candida parapsilosis demonstrates the role of secreted lipase in virulence

Targeted gene deletion in Candida parapsilosis demonstrates the role of secreted lipase in virulence

Candida parapsilosis, an Emerging Fungal Pathogen

Candida parapsilosis Secreted Lipase as an Important Virulence Factor

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