ArticleNucleic acids research2014
Conditional DNA repair mutants enable highly precise genome engineering.
Article in Nucleic acids research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 57 citations in OpenAlex.
- Accelerated Adaptive Evolution of Gram-Negative Bacteria With a Broad-Host-Range Dual Base Editor.Microbial biotechnology · 2026Article
- Continuous multiplexed phage genome editing using recombitrons.Nature biotechnology · 2025Article
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- Recent advances in genome-scale engineering inEngineering microbiology · 2024Review
- Generating Single Nucleotide Point Mutations in E. coli with the No-SCAR System.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Efficient and iterative retron-mediatedSynthetic biology (Oxford, England) · 2022Article
- ssDNA recombineering boosts in vivo evolution of nanobodies displayed on bacterial surfaces.Communications biology · 2021Article
- Recombineering and MAGE.Nature reviews. Methods primers · 2021Article
- Exploring the fitness benefits of genome reduction in Escherichia coli by a selection-driven approach.Scientific reports · 2020Article
- High-Efficiency Multi-site Genomic Editing of Pseudomonas putida through Thermoinducible ssDNA Recombineering.iScience · 2020Article
- CRISPR/Cas9 recombineering-mediated deep mutational scanning of essential genes in Escherichia coli.Molecular systems biology · 2020Article
- A versatile platform strain for high-fidelity multiplex genome editing.Nucleic acids research · 2019Article
- Efficient engineering of chromosomal ribosome binding site libraries in mismatch repair proficient Escherichia coli.Scientific reports · 2017Article
- Enhancing multiplex genome editing by natural transformation (MuGENT) via inactivation of ssDNA exonucleases.Nucleic acids research · 2017Article
- Targeted mutagenesis: A sniper-like diversity generator in microbial engineering.Synthetic and systems biotechnology · 2017Review
- A 'Semi-Protected Oligonucleotide Recombination' Assay for DNA Mismatch Repair in vivo Suggests Different Modes of Repair for Lagging Strand Mismatches.Nucleic acids research · 2017Article
- Systematic genetic dissection of PTS in Vibrio cholerae uncovers a novel glucose transporter and a limited role for PTS during infection of a mammalian host.Molecular microbiology · 2017Article
- RNA Structural Determinants of Optimal Codons Revealed by MAGE-Seq.Cell systems · 2016Article
- Indispensability of Horizontally Transferred Genes and Its Impact on Bacterial Genome Streamlining.Molecular biology and evolution · 2016Article
- LNA modification of single-stranded DNA oligonucleotides allows subtle gene modification in mismatch-repair-proficient cells.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
Abstract
Oligonucleotide-mediated multiplex genome engineering is an important tool for bacterial genome editing. The efficient application of this technique requires the inactivation of the endogenous methyl-directed mismatch repair system that in turn leads to a drastically elevated genomic mutation rate and the consequent accumulation of undesired off-target mutations. Here, we present a novel strategy for mismatch repair evasion using temperature-sensitive DNA repair mutants and temporal inactivation of the mismatch repair protein complex in Escherichia coli. Our method relies on the transient suppression of DNA repair during mismatch carrying oligonucleotide integration. Using temperature-sensitive control of methyl-directed mismatch repair protein activity during multiplex genome engineering, we reduced the number of off-target mutations by 85%, concurrently maintaining highly efficient and unbiased allelic replacement.
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Registered trials
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.