Functional Reconstitution of a Fungal Natural Product Gene Cluster by Advanced Genome Editing

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• 作者：Jakob Weber ; Vito Valiante ; Christina S. Nødvig ; Derek J. Mattern ; Rebecca A. Slotkowski ; Uffe H. Mortensen ; Axel A. Brakhage
• 刊名：ACS Synthetic Biology
• 出版年：2017
• 出版时间：January 20, 2017
• 年：2017
• 卷：6
• 期：1
• 页码：62-68
• 全文大小：347K
• ISSN：2161-5063

文摘

Filamentous fungi produce varieties of natural products even in a strain dependent manner. However, the genetic basis of chemical speciation between strains is still widely unknown. One example is trypacidin, a natural product of the opportunistic human pathogen Aspergillus fumigatus, which is not produced among different isolates. Combining computational analysis with targeted gene editing, we could link a single nucleotide insertion in the polyketide synthase of the trypacidin biosynthetic pathway and reconstitute its production in a nonproducing strain. Thus, we present a CRISPR/Cas9-based tool for advanced molecular genetic studies in filamentous fungi, exploiting selectable markers separated from the edited locus.