ArticleNature protocols2016
Targeted isolation and cloning of 100-kb microbial genomic sequences by Cas9-assisted targeting of chromosome segments.
Article in Nature protocols, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 68 citations in OpenAlex.
- 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
- Wolbachia enhances ovarian development in the rice planthopper Laodelphax striatellus through elevated energy production.Nature communications · 2025Article
- Enhancing Secondary Metabolite Production in Actinobacteria Through Over-Expression of a Medium-Sized SARP Regulator.International journal of molecular sciences · 2025Article
- The landscape of bacterial contractile injection systems across large-scale metagenomes.Microbiology spectrum · 2025Article
- Leveraging the power of long reads for targeted sequencing.Genome research · 2024Review
- A CRISPR-Cas9-Mediated Large-Fragment Assembly Method for Cloning Genomes and Biosynthetic Gene Cluster.Microorganisms · 2024Article
- Recent advances in the direct cloning of large natural product biosynthetic gene clusters.Engineering microbiology · 2023Review
- Applications of Programmable Endonucleases in Sequence- and Ligation-Independent Seamless DNA Assembly.Biomolecules · 2023Review
- Identification and heterologous expression of the globomycin biosynthetic gene cluster.Synthetic and systems biotechnology · 2023Article
- Precise DNA cleavage using CRISPR-SpRYgests.Nature biotechnology · 2023Article
- Breakthrough in efficient cloning and activation of large cryptic biosynthetic gene clusters from high GC actinobacteria.Synthetic and systems biotechnology · 2022Article
- The TelN/tos-assisted precise targeting of chromosome segments (TAPE).Journal of advanced research · 2022Article
- Striving for sustainable biosynthesis: discovery, diversification, and production of antimicrobial drugs in Escherichia coli.Biochemical Society transactions · 2022Review
- Genetic and Biochemical Characterization of Halogenation and Drug Transportation Genes Encoded in the Albofungin Biosynthetic Gene Cluster.Applied and environmental microbiology · 2022Article
- β-Hydroxylation of α-amino-β-hydroxylbutanoyl-glycyluridine catalyzed by a nonheme hydroxylase ensures the maturation of caprazamycin.Communications chemistry · 2022Article
- Multiplying the heterologous production of spinosad through tandem amplification of its biosynthetic gene cluster in Streptomyces coelicolor.Microbial biotechnology · 2022Article
- Activating cryptic biosynthetic gene cluster through a CRISPR-Cas12a-mediated direct cloning approach.Nucleic acids research · 2022Article
- Synthetic Biology Advanced Natural Product Discovery.Metabolites · 2021Review
- Biosynthesis and Heterologous Expression of Cacaoidin, the First Member of the Lanthidin Family of RiPPs.Antibiotics (Basel, Switzerland) · 2021Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cloning of long microbial genomic sequences is an essential tool in synthetic biology and genome engineering. Such long sequences are often difficult to obtain directly by traditional PCR or restriction enzyme digestion, and therefore the cloning of these sequences has remained a technical obstacle in molecular biology. Based on the in vitro application of RNA-guided Cas9 nuclease, the method of Cas9-assisted targeting of chromosome segments (CATCH) cleaves target DNA in vitro from intact bacterial chromosomes embedded in agarose plugs, which can be subsequently ligated with cloning vector through Gibson assembly. Here we describe an optimized protocol of CATCH cloning for the targeted cloning of long genomic sequences of up to 100 kb from microorganisms. The protocol uses standard laboratory equipment and takes ∼8 h of bench time over several days, and it may potentially simplify and accelerate efforts to isolate and clone large gene clusters from microorganisms.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.