ArticleNucleic acids research2019
A versatile platform strain for high-fidelity multiplex genome editing.
Article in Nucleic acids research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Describing the persistence landscape for introducing microbes into complex communities.Applied and environmental microbiology · 2026Review
- An optimized CRISPR/Cas9-based genome editing platform enhances bacteriocin production in Lactiplantibacillus plantarum.Journal of biological engineering · 2026Article
- Cross-order detection of bacteriophage transduction in microbial communities using RNA barcoding.Nature communications · 2026Article
- Identification of proteins influencing CRISPR-associated transposases for enhanced genome editing.Science advances · 2026Article
- Phage-based delivery of CRISPR-associated transposases for targeted bacterial editing.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Coselection of BAC for Escherichia coli chromosomal DNA multiplex automated genome engineering.Biotechnology letters · 2024Article
- Soil microbiome engineering for sustainability in a changing environment.Nature biotechnology · 2023Review
- Retron-mediated multiplex genome editing and continuous evolution in Escherichia coli.Nucleic acids research · 2023Article
- Efficient plasmid transfer via natural competence in a microbial co-culture.Molecular systems biology · 2023Article
- Species- and site-specific genome editing in complex bacterial communities.Nature microbiology · 2022Article
- A standardized genome architecture for bacterial synthetic biology (SEGA).Nature communications · 2021Article
- Synthetic chimeric nucleases function for efficient genome editing.Nature communications · 2019Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
Precision genome editing accelerates the discovery of the genetic determinants of phenotype and the engineering of novel behaviors in organisms. Advances in DNA synthesis and recombineering have enabled high-throughput engineering of genetic circuits and biosynthetic pathways via directed mutagenesis of bacterial chromosomes. However, the highest recombination efficiencies have to date been reported in persistent mutator strains, which suffer from reduced genomic fidelity. The absence of inducible transcriptional regulators in these strains also prevents concurrent control of genome engineering tools and engineered functions. Here, we introduce a new recombineering platform strain, BioDesignER, which incorporates (i) a refactored λ-Red recombination system that reduces toxicity and accelerates multi-cycle recombination, (ii) genetic modifications that boost recombination efficiency, and (iii) four independent inducible regulators to control engineered functions. These modifications resulted in single-cycle recombineering efficiencies of up to 25% with a 7-fold increase in recombineering fidelity compared to the widely used recombineering strain EcNR2. To facilitate genome engineering in BioDesignER, we have curated eight context--neutral genomic loci, termed Safe Sites, for stable gene expression and consistent recombination efficiency. BioDesignER is a platform to develop and optimize engineered cellular functions and can serve as a model to implement comparable recombination and regulatory systems in other bacteria.
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Registered trials
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