Filamentous fungi are historically known to be a rich reservoir of bioactive compounds that are applied in a myriad of fields ranging from crop protection to medicine. The surge of genomic data available shows that fungi remain an excellent source for new pharmaceuticals. However, most of the responsible biosynthetic gene clusters are transcriptionally silent under laboratory growth conditions. Therefore, generic strategies for activation of these clusters are required. Here, we present a genome-editing-free, transcriptional regulation tool for filamentous fungi, based on the CRISPR activation (CRISPRa) methodology.
- Discuss the generation of a CRISPRa / dCas9-VPR system for transcriptional activation
- Describe advantages and disadvantages of Synthetic Transcription Factors
- Discuss modular cloning-based synthetic transcription unit construction