ArticleNucleic acids research2022
Activating cryptic biosynthetic gene cluster through a CRISPR-Cas12a-mediated direct cloning approach.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed.
- Advances in large DNA fragment assembly for microbial cell factory engineering.Quantitative biology (Beijing, China) · 2026Review
- Heterologous expression of NRPS and PKS pathways in Escherichia coli: from bottlenecks to biosynthetic platforms.Essays in biochemistry · 2026Review
- A guide to Direct Pathway Cloning (DiPaC) for natural product discovery.Essays in biochemistry · 2026Review
- Pseudotetraivprolides from Pseudomonas entomophila Provide Insights into the Biosynthesis of Detoxin/Rimosamide-Like Anti-Antibiotics.Angewandte Chemie (International ed. in English) · 2026Article
- Osmotically Tunable Microdroplets Enable Amplification-Free CRISPR Detection of Gene Doping.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Model-Guided Systematic Metabolic Engineering for Enhanced Spinosad Biosynthesis in Saccharopolyspora spinosa NHF132.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Construction and Diversification of Natural Product Biosynthetic Gene Clusters at High Efficiency and Accuracy.ACS synthetic biology · 2025Article
- Analyzing the Challenges and Opportunities Associated With Harnessing New Antibiotics From the Fungal Microbiome.MicrobiologyOpen · 2025Review
- Efficient mining of natural products from Mangrove streptomyces using NP-PRESS.Archives of microbiology · 2025Article
- Overproduction of endusamycin inSynthetic and systems biotechnology · 2025Article
- Synthetic Biology in Natural Product Biosynthesis.Chemical reviews · 2025Review
- Harnessing the Streptomyces-originating type I-E CRISPR/Cas system for efficient genome editing in Streptomyces.Science China. Life sciences · 2025Article
- Self-resistance-gene-guided, high-throughput automated genome mining of bioactive natural products from Streptomyces.Cell systems · 2025Article
- Biology and applications of CRISPR-Cas12 and transposon-associated homologs.Nature biotechnology · 2024Review
- Repurposing endogenous type I-E CRISPR-Cas systems for natural product discovery in Streptomyces.Nature communications · 2024Article
- CRISETR: an efficient technology for multiplexed refactoring of biosynthetic gene clusters.Nucleic acids research · 2024Article
- Mining microbial and metabolic dark matter in extreme environments: a roadmap for harnessing the power of multi-omics data.Advanced biotechnology · 2024Review
- A CRISPR-Cas9-Mediated Large-Fragment Assembly Method for Cloning Genomes and Biosynthetic Gene Cluster.Microorganisms · 2024Article
- CRISPR-aided genome engineering for secondary metabolite biosynthesis in Streptomyces.Journal of industrial microbiology & biotechnology · 2024Review
- Exploring a general multi-pronged activation strategy for natural product discovery in Actinomycetes.Communications biology · 2024Article
Corrections and comments
- Erratum issued
Authors and funding
18 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Direct cloning of biosynthetic gene clusters (BGCs) from microbial genomes facilitates natural product-based drug discovery. Here, by combining Cas12a and the advanced features of bacterial artificial chromosome library construction, we developed a fast yet efficient in vitro platform for directly capturing large BGCs, named CAT-FISHING (CRISPR/Cas12a-mediated fast direct biosynthetic gene cluster cloning). As demonstrations, several large BGCs from different actinomycetal genomic DNA samples were efficiently captured by CAT-FISHING, the largest of which was 145 kb with 75% GC content. Furthermore, the directly cloned, 110 kb long, cryptic polyketide encoding BGC from Micromonospora sp. 181 was then heterologously expressed in a Streptomyces chassis. It turned out to be a new macrolactam compound, marinolactam A, which showed promising anticancer activity. Our results indicate that CAT-FISHING is a powerful method for complicated BGC cloning, and we believe that it would be an important asset to the entire community of natural product-based drug discovery.
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