Examination of Cyp51A-Mediated Azole Resistance in Aspergillus lentulus Using CRISPR/Cas9 Genome Editing

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  • Tateno Madoka
    Division of Infection Control Sciences, Department of Clinical Pharmacy, School of Pharmacy, Showa University Department of Fungal Infection, National Institute of Infectious Diseases
  • Umeyama Takashi
    Department of Fungal Infection, National Institute of Infectious Diseases
  • Inukai Tatsuya
    Department of Fungal Infection, National Institute of Infectious Diseases Department of Microbiology, Tokyo Medical University
  • Takatsuka Shogo
    Department of Fungal Infection, National Institute of Infectious Diseases
  • Hoshino Yasutaka
    Department of Fungal Infection, National Institute of Infectious Diseases
  • Yamagoe Satoshi
    Department of Fungal Infection, National Institute of Infectious Diseases
  • Yamagata Murayama Somay
    Department of Fungal Infection, National Institute of Infectious Diseases
  • Ishino Keiko
    Division of Infection Control Sciences, Department of Clinical Pharmacy, School of Pharmacy, Showa University
  • Miyazaki Yoshitsugu
    Department of Fungal Infection, National Institute of Infectious Diseases

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  • Examination of Cyp51A-Mediated Azole Resistance in <i>Aspergillus lentulus</i> Using CRISPR/Cas9 Genome Editing

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Aspergillus lentulus was first reported in 2005 as a cryptic species of Aspergillus fumigatus, and since then, its resistance to azole drugs and the high mortality rate of infected individuals have emerged as problems. Although it has been reported that P450 14-α sterol demethylase (Cyp51) is involved in azole resistance in A. lentulus, the specific resistance mechanism has not been elucidated. In this study, we successfully introduced the entire A. fumigatus cyp51A gene into the cyp51A locus in A. lentulus using the CRISPR/Cas9 genome-editing system. The A. lentulus strains harboring A. fumigatus cyp51A showed reduced minimum inhibitory concentrations for itraconazole and voriconazole compared with those of the parent strain. This finding suggests that Cyp51A is involved in azole resistance in A. lentulus and may contribute to the elucidation of the mechanism of resistance to azole drugs via Cyp51A and to the development of new antifungal drugs. In addition, our successful application of the CRISPR/Cas9 system to A. lentulus opens the door to examination of other gene functions in this fungus.

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