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Navarro Ros, Eusebio

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Navarro Ros, Eusebio
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Universidad de Murcia. Departamento de Genética y Microbiología
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    Stable and reproducible homologous recombination enables CRISPR-based engineering in the fungus Rhizopus microsporus
    (Cell Press, 2021-12-20) Lax Molina, Carlos; Navarro Mendoza, María Isabel; Pérez Arques, Carlos; Navarro Ros, Eusebio; Nicolás Molina, Francisco Esteban; Garre Mula, Victoriano; Genética y Microbiología; Facultades de la UMU::Facultad de Biología
    Mucormycosis is a lethal and emerging disease that has lacked a genetic model fulfilling both high virulence and the possibility of performing stable and reproducible gene manipulation by homologous recombination (HR). Here, we developed a new methodology to successfully perform HR in Rhizopus microsporus. We isolated an uracil auxotrophic recipient strain and optimized the critical steps in the genetic transformation of this fungus. This was followed by an adaptation of a plasmid-free CRISPR-Cas9 system coupled with microhomology repair templates. We reproducibly generated stable mutants in the genes leuA and crgA, encoding a 3-isopropylmalate dehydratase and an ubiquitin ligase, respectively. Our new genetic model showed that mutations in the gene pyrF, a key virulence gene in several bacterial and fungal pathogens, correlated with an avirulent phenotype in an immunocompetent murine host. This was reverted by gene complementation, showing the broad possibilities of our methodology.
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    A ribonuclease III involved in virulence of Mucorales fungi has evolved to cut exclusively single-stranded RNA
    (2021) Falk, Sebastian; Zapata Pérez, Rubén; Sánchez Ferrer, Álvaro; Nicolás Molina, Francisco Esteban; Sánchez Ferrer, Álvaro; Navarro Ros, Eusebio; Garre Mula, Victoriano; Padmanabhan Iyer, Subramanian; Cánovas Márquez, José Tomás; Genética y Microbiología
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