Journal
GENES
Volume 9, Issue 12, Pages -Publisher
MDPI
DOI: 10.3390/genes9120613
Keywords
Mucor circinelloides; mucormycosis; firefly luciferase; reporter strain; bioluminescence
Categories
Funding
- EMBO short-term fellowship [6856]
- Christian Doppler Forschungsgesellschaft (CD-Labor Invasive Pilzinfektionen)
- Ministerio de Educacion, Cultura y Deporte, Spain [FPU-14/01832]
- Ministerio de Economia y Competitividad, Spain [RYC-2014-15844]
- Ministerio de Economia y Competitividad, Spain (FEDER) [BFU2015-65501-P]
- Fundacion Seneca-Agencia de Ciencia y Tecnologia de la Region de Murcia, Spain [19339/PI/14]
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Invasive fungal infections caused by Mucorales (mucormycosis) have increased worldwide. These life-threatening infections affect mainly, but not exclusively, immunocompromised patients, and are characterized by rapid progression, severe tissue damage and an unacceptably high rate of mortality. Still, little is known about this disease and its successful therapy. New tools to understand mucormycosis and a screening method for novel antimycotics are required. Bioluminescent imaging is a powerful tool for in vitro and in vivo approaches. Hence, the objective of this work was to generate and functionally analyze bioluminescent reporter strains of Mucor circinelloides, one mucormycosis-causing pathogen. Reporter strains were constructed by targeted integration of the firefly luciferase gene under control of the M. circinelloides promoter Pzrt1. The luciferase gene was sufficiently expressed, and light emission was detected under several conditions. Phenotypic characteristics, virulence potential and antifungal susceptibility were indifferent to the wild-type strains. Light intensity was dependent on growth conditions and biomass, being suitable to determine antifungal efficacy in vitro. This work describes for the first time the generation of reporter strains in a basal fungus that will allow real-time, non-invasive infection monitoring in insect and murine models, and the testing of antifungal efficacy by means other than survival.
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