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.
| Original language | English |
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| Article number | 134326 |
| Number of pages | 8 |
| Journal | International Journal of Biological Macromolecules |
| Volume | 277 |
| Issue number | Pt 2 |
| Early online date | 30 Jul 2024 |
| DOIs | |
| Publication status | Published - Oct 2024 |
Bibliographical note
Publisher Copyright:© 2024 The Authors
Funding
XC is supported by a grant of the China Scholarship Council CSC. During the preparation of this work the authors used ChatGPT in order to improve language. After using this tool, the authors reviewed and edited the content as needed and take full responsibility for the content of the publication.
| Funders |
|---|
| China Scholarship Council |
Keywords
- Aspergillus
- Epigenetic modification
- Fungus