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Future Microbiol ; 14: 1383-1396, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31659913

RESUMO

Aim: In this study, the CRISPR gene-editing approach was used to simultaneously inactivate all three members of the ALS gene family in the opportunistic pathogen Candida orthopsilosis. Materials & methods: Using a single gRNA and repair template, CRISPR-edited clones were successfully generated in a one-step process in both C. orthopsilosis reference and clinical strains. Results: The phenotypic characterization of the ALS triple-edited strains revealed no impact on growth in liquid or solid media. However, pseudohyphal formation and the ability to adhere to human buccal epithelial cells were significantly decreased in triple-edited clones. Conclusion: Our CRISPR/Cas9 system is a powerful tool for simultaneous editing of fungal gene families, which greatly accelerates the generation of multiple gene-edited Candida strains. Data deposition: Nucleotide sequence data are available in the GenBank databases under the accession numbers MK875971, MK875972, MK875973, MK875974, MK875975, MK875976, MK875977.


Assuntos
Proteína 9 Associada à CRISPR/genética , Sistemas CRISPR-Cas , Candida parapsilosis/genética , Edição de Genes/métodos , Genes Fúngicos , Sequência de Bases , Candida parapsilosis/crescimento & desenvolvimento , Candidíase/microbiologia , Adesão Celular , Células Cultivadas , Células Epiteliais/microbiologia , Humanos , Hifas/crescimento & desenvolvimento , Boca/citologia , Família Multigênica , RNA Guia de Cinetoplastídeos/genética
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