ArticleJournal of molecular biology2011
Probing cellular processes with oligo-mediated recombination and using the knowledge gained to optimize recombineering.
Article in Journal of molecular biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 95 papers.
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Who cites it
95 citing papers in PubMed, 167 citations in OpenAlex.
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- A genetic manipulation tool based on the GP35 recombinase for targeted gene editing in mycoplasmas of ruminants.The Journal of biological chemistry · 2026Article
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- Genome-Editing Tools for Lactic Acid Bacteria: Past Achievements, Current Platforms, and Future Directions.International journal of molecular sciences · 2025Review
- Genome editing outcomes reveal mycobacterial NucS participates in a short-patch repair of DNA mismatches.Nucleic acids research · 2024Article
- Development of a Recombineering System for the AcetogenACS synthetic biology · 2024Article
- Recent advances in genome-scale engineering inEngineering microbiology · 2024Review
- Genome Editing Outcomes Reveal Mycobacterial NucS Participates in a Short-Patch Repair of DNA Mismatches.bioRxiv : the preprint server for biology · 2023Article
- Retron-mediated multiplex genome editing and continuous evolution in Escherichia coli.Nucleic acids research · 2023Article
- CRISPR/Cas9-assisted ssDNA recombineering for site-directed mutagenesis and saturation mutagenesis of plasmid-encoded genes.Biotechnology letters · 2023Article
- Recombineering: Genetic Engineering in Escherichia coli Using Homologous Recombination.Current protocols · 2023Article
- Single-nucleotide resolution detection of Topo IV cleavage activity in theFrontiers in microbiology · 2023Article
- Synthetic biology tools for engineeringComputational and structural biotechnology journal · 2023Review
- A study on L-threonine and L-serine uptake inFrontiers in microbiology · 2023Article
- Article
- Recombineering in Non-Model Bacteria.Current protocols · 2022Article
- Phenotype of Mrps5-Associated Phylogenetic Polymorphisms Is Intimately Linked to Mitoribosomal Misreading.International journal of molecular sciences · 2022Article
- Resolving the structure of phage-bacteria interactions in the context of natural diversity.Nature communications · 2022Article
- Efficient and iterative retron-mediatedSynthetic biology (Oxford, England) · 2022Article
- Recombineering in Staphylococcus aureus.Methods in molecular biology (Clifton, N.J.) · 2022Article
35 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors at 3 institutions in 1 country.
Funding
Abstract
Recombination with single-strand DNA oligonucleotides (oligos) in Escherichia coli is an efficient and rapid way to modify replicons in vivo. The generation of nucleotide alteration by oligo recombination provides novel assays for studying cellular processes. Single-strand exonucleases inhibit oligo recombination, and recombination is increased by mutating all four known exonucleases. Increasing oligo concentration or adding nonspecific carrier oligo titrates out the exonucleases. In a model for oligo recombination, λ Beta protein anneals the oligo to complementary single-strand DNA at the replication fork. Mismatches are created, and the methyl-directed mismatch repair (MMR) system acts to eliminate the mismatches inhibiting recombination. Three ways to evade MMR through oligo design include, in addition to the desired change (1) a C·C mismatch 6 bp from that change; (2) four or more adjacent mismatches; or (3) mismatches at four or more consecutive wobble positions. The latter proves useful for making high-frequency changes that alter only the target amino acid sequence and even allows modification of essential genes. Efficient uptake of DNA is important for oligo-mediated recombination. Uptake of oligos or plasmids is dependent on media and is 10,000-fold reduced for cells grown in minimal versus rich medium. Genomewide engineering technologies utilizing recombineering will benefit from both optimized recombination frequencies and a greater understanding of how biological processes such as DNA replication and cell division impact recombinants formed at multiple chromosomal loci. Recombination events at multiple loci in individual cells are described here.
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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.