ArticleACS synthetic biology2025
Construction and Diversification of Natural Product Biosynthetic Gene Clusters at High Efficiency and Accuracy.
Article in ACS synthetic biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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Who cites it
3 citing papers in PubMed.
- Biosynthetic gene clusters inmSystems · 2026Article
- Inducible CRISPRi enables efficient and high-fidelity genome editing in Streptomyces.Nucleic acids research · 2026Article
- γ-Butyrolactone-induced coelimycin synthesis inhibits AtrA-dependent actinorhodin overproduction inmBio · 2026Article
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Authors and funding
3 authors.
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Abstract
Biosynthetic gene clusters (BGCs) encode the biosynthesis of natural products, which serve as the foundation for therapeutics such as antibiotics, anticancer agents, antifungals, and immunosuppressants. The vast majority of the BGCs remain uncharacterized due to lack of expression or inability to cultivate the native host, making refactoring and expression of BGCs in optimized hosts a prerequisite for genome-based drug discovery. Transformation-associated recombination (TAR) cloning and Gibson assembly are error prone due to the use of homologous recombination. Here, we present a BGC cloning and refactoring strategy based on a hierarchical Golden Gate Assembly (GGA), which enables systematic pathway engineering and mutagenesis with unprecedented accuracy and efficiency. We constructed the 23 kb actinorhodin (ACT) BGC and 23 mutant derivatives with either one of the
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