Functional analysis of FlbA-regulated transcription factor genes in Aspergillus niger using a multiplexed CRISPRoff system.
Int J Biol Macromol
; 277(Pt 2): 134326, 2024 Oct.
Article
in En
| MEDLINE
| ID: mdl-39089555
ABSTRACT
FlbA of Aspergillus niger (indirectly) regulates 36 transcription factor (TF) genes. As a result, it promotes sporulation and represses vegetative growth, protein secretion and lysis. In this study, the functions of part of the FlbA-regulated TF genes were studied by using CRISPRoff. This system was recently introduced as an epigenetic tool for modulating gene expression in A. niger. A plasmid encompassing an optimized CRISPRoff system as well as a library of sgRNA genes that target the promoters of the 36 FlbA-regulated TF genes was introduced in A. niger. Out of 24 transformants that exhibited a sporulation phenotype, 12 and 18 strains also showed a biomass and secretion phenotype, respectively. The transforming sgRNAs, and thus the genes responsible for the phenotypes, were identified from five of the transformants. The results show that the genes dofA, dofB, dofC, dofD, and socA are involved in sporulation and extracellular enzyme activity, while dofA and socA also play roles in biomass formation. Overall, this study shows that the multiplexed CRISPRoff system can be effectively used for functional analysis of genes in a fungus.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Aspergillus niger
/
Transcription Factors
/
Fungal Proteins
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Gene Expression Regulation, Fungal
Language:
En
Journal:
Int J Biol Macromol
Year:
2024
Document type:
Article
Country of publication: