CRISPR-Mediated Activation of Biosynthetic Gene Clusters for Bioactive Molecule Discovery in Filamentous Fungi
نویسندگان
چکیده
منابع مشابه
Metabolic pathway gene clusters in filamentous fungi.
Clusters of functionally related genes are a general feature of prokaryotic gene organization but are much less prevalent in eukaryotes. The discovery that genes for certain types of metabolic pathways are clustered in filamentous fungi is relatively recent. Only 8 years ago in a discussion of the proline utilization pathway genes, Hull et al. (1989) commented that ‘‘The organization and regula...
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Filamentous fungi are a large group of diverse and economically important microorganisms. Large-scale gene disruption strategies developed in budding yeast are not applicable to these organisms because of their larger genomes and lower rate of targeted integration (TI) during transformation. We developed transposon-arrayed gene knockouts (TAGKO) to discover genes and simultaneously create gene ...
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Here we report an efficient CRISPR-Cas9 knock-in strategy to activate silent biosynthetic gene clusters (BGCs) in streptomycetes. We applied this one-step strategy to activate multiple BGCs of different classes in five Streptomyces species and triggered the production of unique metabolites, including a novel pentangular type II polyketide in Streptomyces viridochromogenes. This potentially scal...
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Biosynthetic pathways of secondary metabolites from fungi are currently subject to an intense effort to elucidate the genetic basis for these compounds due to their large potential within pharmaceutics and synthetic biochemistry. The preferred method is methodical gene deletions to identify supporting enzymes for key synthases one cluster at a time. In this study, we design and apply a DNA expr...
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The use of DNA sequencing to guide the discovery of natural products has emerged as a new paradigm for revealing chemistries encoded in bacterial genomes. A major obstacle to implementing this approach to natural product discovery is the transcriptional silence of biosynthetic gene clusters under laboratory growth conditions. Here we describe an improved yeast-based promoter engineering platfor...
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ژورنال
عنوان ژورنال: ACS Synthetic Biology
سال: 2020
ISSN: 2161-5063,2161-5063
DOI: 10.1021/acssynbio.0c00197