ArticleNature communications2021
Cas12a-assisted precise targeted cloning using in vivo Cre-lox recombination.
Article in Nature communications, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 62 papers, 1 of them a synthesis that pooled it.
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Who cites it
62 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Advancements in the Application of Ribosomally Synthesized and Post-Translationally Modified Peptides (RiPPs).Biomolecules · 2024Pooled it
- The PreKit platform: Cryptic gene clusters activation and high-titer compound production.Synthetic and systems biotechnology · 2026Article
- Antibiotic-producing microorganisms: taxonomic diversity, screening strategies, and emerging and underexploited antibacterial targets.Archives of microbiology · 2026Review
- Accelerating natural product discovery, characterization and engineering by biofoundries.Natural product reports · 2026Review
- The FAD-Dependent Monooxygenase Albo65 Mediates Self-Resistance in Albofungin Biosynthesis.JACS Au · 2026Article
- Heterologous expression of NRPS and PKS pathways in Escherichia coli: from bottlenecks to biosynthetic platforms.Essays in biochemistry · 2026Review
- Reimagining Lignin Valorization: Synthetic Biology-Enabled Sustainable Aromatic Carbon Biomanufacturing.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Review
- DiscERN: an automated genome mining tool for the discovery of evolutionarily related natural products.Nature communications · 2026Article
- Heterologous production of a plant biostimulant in Streptomyces albidoflavus.Applied microbiology and biotechnology · 2026Article
- Engineering an IndustrialMicroorganisms · 2026Article
- Indigomycin, a new pyridinetrione from Streptomyces albus J1074 harboring the indigoidine biosynthetic gene cluster.The Journal of antibiotics · 2026Article
- Article
- Strategies to decipher silent biosynthetic gene clusters in actinomycetes.Archives of microbiology · 2025Review
- A Versatile Enzymatic Pathway for Modification of Peptide C‑Termini.ACS central science · 2025Article
- Construction and Diversification of Natural Product Biosynthetic Gene Clusters at High Efficiency and Accuracy.ACS synthetic biology · 2025Article
- StreptoCAD: An Open-Source Software Toolbox Automating Genome Engineering Workflows in Streptomycetes.ACS synthetic biology · 2025Article
- The Role of Nucleotide Sequencing in Natural Product Drug Discovery.Journal of natural products · 2025Review
- Brazilian Biomes as Promising Resources of Rhodotorula Yeasts for the Biotechnological Production of Carotenoids.Chemistry & biodiversity · 2025Review
- CRISPR/Cas12a-Based Biosensing: Advances in Mechanisms and Applications for Nucleic Acid Detection.Biosensors · 2025Review
- Overproduction of endusamycin inSynthetic and systems biotechnology · 2025Article
2 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Direct cloning represents the most efficient strategy to access the vast number of uncharacterized natural product biosynthetic gene clusters (BGCs) for the discovery of novel bioactive compounds. However, due to their large size, repetitive nature, or high GC-content, large-scale cloning of these BGCs remains an overwhelming challenge. Here, we report a scalable direct cloning method named Cas12a-assisted precise targeted cloning using in vivo Cre-lox recombination (CAPTURE) which consists of Cas12a digestion, a DNA assembly approach termed T4 polymerase exo + fill-in DNA assembly, and Cre-lox in vivo DNA circularization. We apply this method to clone 47 BGCs ranging from 10 to 113 kb from both Actinomycetes and Bacilli with ~100% efficiency. Heterologous expression of cloned BGCs leads to the discovery of 15 previously uncharacterized natural products including six cyclic head-to-tail heterodimers with a unique 5/6/6/6/5 pentacyclic carbon skeleton, designated as bipentaromycins A-F. Four of the bipentaromycins show strong antimicrobial activity to both Gram-positive and Gram-negative bacteria such as methicillin-resistant Staphylococcus aureus, vancomycinresistant Enterococcus faecium, and bioweapon Bacillus anthracis. Due to its robustness and efficiency, our direct cloning method coupled with heterologous expression provides an effective strategy for large-scale discovery of novel natural products.
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