ArticleMicrobial cell factories2020
Homology-dependent recombination of large synthetic pathways into E. coli genome via λ-Red and CRISPR/Cas9 dependent selection methodology.
Article in Microbial cell factories, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- CRISPR-Cas-Mediated Reprogramming Strategies to Overcome Antimicrobial Resistance.Pharmaceutics · 2026Review
- Development and Optimization of CRISPR/Cas9-Assisted Recombineering inmicroPublication biology · 2026Article
- Site-Directed Genome Integration via Recombinase-Mediated Cassette Exchange (RMCE) inACS synthetic biology · 2025Article
- Review
- CRISPR-Cas9-based genome-editing technologies in engineering bacteria for the production of plant-derived terpenoids.Engineering microbiology · 2024Review
- The Influence of Homologous Arm Length on Homologous Recombination Gene Editing Efficiency Mediated by SSB/CRISPR-Cas9 inMicroorganisms · 2024Article
- An adapted method for Cas9-mediated editing reveals the species-specific role of β-glucoside utilization driving competition betweenJournal of bacteriology · 2024Article
- Advances and applications of CRISPR/Cas-mediated interference inEngineering microbiology · 2024Review
- The non-mevalonate pathway requires a delicate balance of intermediates to maximize terpene production.Applied microbiology and biotechnology · 2024Article
- OptimizingFrontiers in microbiology · 2024Review
- The expanded CRISPR toolbox for constructing microbial cell factories.Trends in biotechnology · 2024Review
- CRISPR-Associated Transposase for Targeted Mutagenesis in Diverse Proteobacteria.ACS synthetic biology · 2023Article
- Mobile and Self-Sustained Data Storage in an Extremophile Genomic DNA.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- Toward bioproduction of oxo chemicals from CBiotechnology for biofuels and bioproducts · 2022Review
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3 authors.
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Abstract
backgroundMetabolic engineering frequently needs genomic integration of many heterologous genes for biosynthetic pathway assembly. Despite great progresses in genome editing for the model microorganism Escherichia coli, the integration of large pathway into genome for stabilized chemical production is still challenging compared with small DNA integration.
resultsWe have developed a λ-Red assisted homology-dependent recombination for large synthetic pathway integration in E. coli. With this approach, we can integrate as large as 12 kb DNA module into the chromosome of E. coli W3110 in a single step. The efficiency of this method can reach 100%, thus markedly improve the integration efficiency and overcome the limitation of the integration size adopted the common method. Furthermore, the limiting step in the methylerythritol 4-phosphate (MEP) pathway and lycopene synthetic pathway were integrated into the W3110 genome using our system. Subsequently, the yields of the final strain were increased 106 and 4.4-fold compared to the initial strain and the reference strain, respectively.
conclusionsIn addition to pre-existing method, our system presents an optional strategy for avoiding using plasmids and a valuable tool for large synthetic pathway assembly in E. coli.
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