ArticleApplied and environmental microbiology2015
Coupling the CRISPR/Cas9 System with Lambda Red Recombineering Enables Simplified Chromosomal Gene Replacement in Escherichia coli.
Article in Applied and environmental microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 128 papers.
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Who cites it
128 citing papers in PubMed.
- One Plasmid Is All You Need: Genome Editing in Escherichia coli Using Endogenous TnpB and Endogenous Recombination System.Biotechnology and bioengineering · 2026Article
- A Lambda Red Recombinase-Assisted CRISPR/Cas9 System for Efficient and Precise Genome Editing in Agrobacterium tumefaciens.Plant biotechnology journal · 2026Article
- Nucleases and Their Inhibitors: Exploring Biological Roles, Industrial Applications, and Challenges in Heterologous Expression.Biotechnology journal · 2026Review
- Chain-Length Regulation by WzzE Is Necessary for, but Genetically Separable From, Cyclic Enterobacterial Common Antigen Synthesis.Molecular microbiology · 2026Article
- Single-Step Chromosomal Engineering Integrates Biocontainment and Therapeutic Function for Regulatory-Oriented Live Biotherapeutic Chassis Design.Pharmaceutics · 2026Article
- Quorum-sensing regulator LsrR modulates resistance to oxidative stress by interfering with sulfate assimilation in avian pathogenicJournal of bacteriology · 2026Article
- Genomically integrated orthogonal translation system in Escherichia coli enables production of functional modified [NiFe]-hydrogenases.Microbial cell factories · 2026Article
- Targeted DNA ADP-ribosylation triggers templated repair in bacteria and base mutagenesis in eukaryotes.Nature biotechnology · 2026Article
- RECC: A Red/ET-CRISPR/Cas9-based system enabling genome mining of marineSynthetic and systems biotechnology · 2026Article
- Genetically Modified Lactic Acid Bacteria in the EU Food Chain: Applications, Benefits, and Risk Assessment.International journal of molecular sciences · 2026Review
- PE-STAR: prime editing with SOS-triggered and RecJ-augmented repair enables high-efficiency editing in Escherichia coli.Nucleic acids research · 2026Article
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- Functional Analysis of CRISPR-Cas9-Mediated Gene Deletion inMicroorganisms · 2026Article
- Development and Optimization of CRISPR/Cas9-Assisted Recombineering inmicroPublication biology · 2026Article
- A comprehensive review of genomic-scale genetic engineering as a strategy to improve bacterial productivity.Microbiology (Reading, England) · 2025Review
- A three-plasmid-containing CRISPR-Cas9 platform to engineerApplied and environmental microbiology · 2025Article
- High-throughput conjugation reveals strain specific recombination patterns enabling precise trait mapping in Escherichia coli.PLoS genetics · 2025Article
- Lethal endotoxin (ccdB) based counterselection improved the efficiency of sequential gene editing in Escherichia coli.Biotechnology letters · 2025Article
- SELECT: high-precision genome editing strategy via integration of CRISPR-Cas and DNA damage response for cross-species applications.Nucleic acids research · 2025Article
- Study on the framework of ATP energy cycle system in Escherichia coli.Applied microbiology and biotechnology · 2025Article
68 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
To date, most genetic engineering approaches coupling the type II Streptococcus pyogenes clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system to lambda Red recombineering have involved minor single nucleotide mutations. Here we show that procedures for carrying out more complex chromosomal gene replacements in Escherichia coli can be substantially enhanced through implementation of CRISPR/Cas9 genome editing. We developed a three-plasmid approach that allows not only highly efficient recombination of short single-stranded oligonucleotides but also replacement of multigene chromosomal stretches of DNA with large PCR products. By systematically challenging the proposed system with respect to the magnitude of chromosomal deletion and size of DNA insertion, we demonstrated DNA deletions of up to 19.4 kb, encompassing 19 nonessential chromosomal genes, and insertion of up to 3 kb of heterologous DNA with recombination efficiencies permitting mutant detection by colony PCR screening. Since CRISPR/Cas9-coupled recombineering does not rely on the use of chromosome-encoded antibiotic resistance, or flippase recombination for antibiotic marker recycling, our approach is simpler, less labor-intensive, and allows efficient production of gene replacement mutants that are both markerless and "scar"-less.
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