Genetic editing using the CRISPR/Cas9 system of the nicotinamide adenine dinucleotide transporter from Aspergillus fumigatus

• Grant number: 18/09958-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2018
• End date: July 31, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal Investigator: Sergio Akira Uyemura
• Grantee: Maycon Marção
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

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. Recently, we have identified and characterized a sequence which encodes the mitochondrial nicotinamine adenine dinucleotide carrier (Ndt1) from A. fumigatus and we conducted the heterologous expression in S. cerevisiae. It expression improved the severe delay in growth of the double mutant Dndt1Dndt2 of the S. cerevisiae in non-fermentable carbon source. In addition, through by proposal 2016/23380-3, we are introducing in the laboratory the gene-editing tool CRISPR/Cas9. The student has been working to get of a mutant that expresses constitutively Cas9 endonuclease and just turn it with gRNA for the ndt1 gene.