Biochemical and Molecular Studies of mitochondrial components from Aspergillus fumigatus fungi: Genic edition using CRISP/Cas9 system of the mitochondrial Mgr2 (Romo1 orthologous).

Abstract

Aspergillus fumigatus is a saprophytic fungus, which has been found in all regions of the World. It is believed that the natural ecological niche is the soil where it grows and survives in decaying plants. In recent years, the fungus A. fumigatus is no longer just a minor saprophytic fungus, to become a major opportunistic pathogen in immunosuppressed patients. In the last 18 years, our laboratory has studied the role of mitochondria and its components in bioenergetics and biology of this fungus. In this sense, the project will study the function of the small subunit of Tim23 complex, the Mgr2 whose ortholog (Romo1) has role in modulation of reactive oxygen species in mammals. At the same time, it will be introduce in the laboratory a new technique using gene-editing tool CRISP/Cas9. The knockout of genes in A. fumigatus is established in the laboratory by gene disruption using PCR-based strategy and in vivo recombination in yeast, but it is not a trivial process. The proposal is to get of a mutant that expresses with Cas9 and just turn it with the plasmid with the target gene. This will create a facility to obtain mutants with silenced genes. Now, there is no research group in Aspergillus in São Paulo State, which is using this technique with support by FAPESP. (AU)