CRISPR/Cas9 technology enables the development of the filamentous ascomycete fungus Penicillium subrubescens as a new industrial enzyme producer.
Penicillium subrubescens is an ascomycete fungus with an enriched content of specific carbohydrate-active enzyme families involved in plant biomass degradation, which makes this strain a promising industrial cell factory for enzyme production. The development of tools that allow genetic manipulation is crucial for further strain improvement and the functional characterization of its genes. In this context, the CRISPR/Cas9 system represents an excellent option for genome editing due to its high efficiency and versatility. To establish CRISPR/Cas9 genome editing in P. subrubescens, first a method for protoplast generation and transformation was developed, using hygromycin as selection marker. Then the CRISPR/Cas9 system was established in P. subrubescens by successfully deleting the ku70 gene, which is involved in the non-homologous end joining DNA repair mechanism. Phenotypic characterization of the mutants showed that ku70 mutation did not affect P. subrubescens growth at optimal temperature and Δku70 strains showed similar protein production pattern to the wild type.
['Salazar-Cerezo|Sonia|S|', 'Kun|Roland S|RS|', 'de Vries|Ronald P|RP|', 'Garrigues|Sandra|S|']
D064113:CRISPR-Cas Systems D004798:Enzymes D000072669:Gene Editing D016681:Genome, Fungal D007218:Industrial Microbiology D010407:Penicillium D010641:Phenotype
CRISPR/Cas9 Cell factory Genome editing ku70 Filamentous fungi Fungal transformation