Related Experiment Video
Updated: Oct 3, 2025
CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans
Published on: November 14, 2018
Examination of Cyp51A-Mediated Azole Resistance in Aspergillus lentulus Using CRISPR/Cas9 Genome Editing
Madoka Tateno1,2, Takashi Umeyama2, Tatsuya Inukai2,3
1Division of Infection Control Sciences, Department of Clinical Pharmacy, School of Pharmacy, Showa University.
Abstract:
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.
Insights
Aspergillus lentulus exhibits azole drug resistance, a significant clinical challenge. This study demonstrates that introducing the Aspergillus fumigatus cyp51A gene reduces azole resistance in A. lentulus, suggesting Cyp51A
Area of Science:
- Mycology
- Antimicrobial Resistance
- Molecular Biology
Background:
- Aspergillus lentulus, a cryptic species of A. fumigatus, presents challenges due to azole drug resistance and high mortality rates.
- The P450 14-α sterol demethylase (Cyp51) enzyme is implicated in azole resistance, but the precise mechanism in A. lentulus remains unclear.
Purpose of the Study:
- To investigate the role of Cyp51A in azole resistance in Aspergillus lentulus.
- To utilize the CRISPR/Cas9 system for genetic manipulation in A. lentulus.
Main Methods:
- CRISPR/Cas9 genome editing was employed to introduce the Aspergillus fumigatus cyp51A gene into the cyp51A locus of A. lentulus.
- Minimum inhibitory concentrations (MICs) for azole drugs were determined for engineered and parent strains.
Main Results:
- A. lentulus strains engineered to express A. fumigatus cyp51A exhibited reduced minimum inhibitory concentrations for itraconazole and voriconazole.
- Successful gene introduction via CRISPR/Cas9 demonstrates its utility for functional genomics in A. lentulus.
Conclusions:
- Cyp51A plays a significant role in azole resistance in Aspergillus lentulus.
- These findings contribute to understanding azole resistance mechanisms and may guide the development of novel antifungal therapies.
- The established CRISPR/Cas9 system facilitates further genetic studies in A. lentulus.
More Related Videos
Related Concept Videos
01:28CRISPR/Cas9 Genome Editing
01:59CRISPR