Abstract
The full potential of fungal secondary metabolism has until recently been impeded by the lack of universal genetic tools for most species. However, the emergence of several CRISPR-Cas9-based genome editing systems adapted for several genera of filamentous fungi have now opened the doors for future efforts in discovery of novel natural products and elucidation and engineering of their biosynthetic pathways in fungi where no genetic tools are in place. So far, most studies have focused on demonstrating the performance of CRISPR-Cas9 in various fungal model species, and recently we presented a versatile CRISPR-Cas9 system that can be successfully applied in several diverse Aspergillus species. Here we take it one step further and show that our system can be used also in a phylogenetically distinct and largely unexplored species from the genus of Talaromyces. Specifically, we exploit CRISPR-Cas9-based genome editing to identify a new gene in T. atroroseus responsible for production of polyketide-nonribosomal peptide hybrid products, hence, linking fungal secondary metabolites to their genetic origin in a species where no genetic engineering has previously been performed.
Highlights
Filamentous fungi are known as prolific producers of numerous industrially important enzymes as well as a diverse spectrum of natural products
First we tested whether the CRISPR-Cas9 system that we have previously developed for Aspergillus could be used directly in T. atroroseus
Amongst the homologous sequences identified in this manner, UA08_00425 was the best match as judged by the size of the ORF and by the high sequence similarities to the corresponding enzymes encoded by wA from A. nidulans (ID: 62%; 99% query coverage) and albA from A. niger (ID: 63%; 99% query coverage)
Summary
Filamentous fungi are known as prolific producers of numerous industrially important enzymes as well as a diverse spectrum of natural products. The latter constitutes an immense reservoir of compounds of biological and medical interest, and many products originating from fungal secondary metabolism are used today in the pharmaceutical industry as e.g. antibiotics, anticancer drugs, cholesterol-lowering agents, and immunosuppressive drugs [1]. The lack of genetic tools available for most fungal species has for many years been the major obstacle for exploring the molecular biology and biochemistry of all but a few model fungi. The enormous increase in sequencing projects over the past years has revealed the PLOS ONE | DOI:10.1371/journal.pone.0169712. The enormous increase in sequencing projects over the past years has revealed the PLOS ONE | DOI:10.1371/journal.pone.0169712 January 5, 2017
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