ABSTRACT
In this study, CRISPR/Cas9 genome editing was used to knockout the bgl2 gene encoding intracellular ß-glucosidase filamentous fungus Penicillium verruculosum. This resulted in a dramatic reduction of secretion of cellulolytic enzymes. The study of P. verruculosum Δbgl2 found that the transcription of the cbh1 gene, which encodes cellobiohydrolase 1, was impaired when induced by cellobiose and cellotriose. However, the transcription of the cbh1 gene remains at level of the host strain when induced by gentiobiose. This implies that gentiobiose is the true inducer of the cellulolytic response in P. verruculosum, in contrast to Neurospora crassa where cellobiose acts as an inducer.
Subject(s)
Penicillium , beta-Glucosidase , Penicillium/genetics , Penicillium/enzymology , beta-Glucosidase/metabolism , beta-Glucosidase/genetics , Gene Expression Regulation, Fungal , Fungal Proteins/genetics , Fungal Proteins/metabolism , Cellulose/metabolism , Cellobiose/metabolism , CRISPR-Cas Systems , Gene Knockout Techniques , Neurospora crassa/genetics , Neurospora crassa/enzymology , Cellulose 1,4-beta-Cellobiosidase/metabolism , Cellulose 1,4-beta-Cellobiosidase/genetics , Gene EditingABSTRACT
The review describes the CRISPR/CAS system and its adaptation for the genome editing in filamentous fungi commonly used for production of enzyme complexes, enzymes, secondary metabolites, and other compounds used in industrial biotechnology and agriculture. In the second part of this review, examples of the CRISPR/CAS technology application for improving properties of the industrial strains of fungi from the Trichoderma, Aspergillus, Penicillium, and other genera are presented. Particular attention is given to the efficiency of genome editing, as well as system optimization for specific industrial producers.
