ArticleNucleic acids research2020
RedEx: a method for seamless DNA insertion and deletion in large multimodular polyketide synthase gene clusters.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed, 39 citations in OpenAlex.
- Combinatorial genome engineering of pseudorabies virus Bartha by developing a reverse genetic system based on three overlapping genomic segments.Journal of advanced research · 2026Article
- Development of efficient genetic toolkits and heterologous chassis for bioprospecting of myxobacteria.Nature communications · 2026Article
- Heterologous production of epothilones inSynthetic and systems biotechnology · 2026Article
- Developing the protein quality control system inSynthetic and systems biotechnology · 2026Article
- Advances in natural product discovery: strategies, technologies, and insights.Natural products and bioprospecting · 2026Review
- iCASRED, a scarless DNA editing tool inSynthetic and systems biotechnology · 2025Article
- Improving polyketide biosynthesis by rescuing the translation of truncated mRNAs into functional polyketide synthase subunits.Nature communications · 2025Article
- Seamless site-directed mutagenesis in complex cloned DNA sequences using the RedEx method.Nature protocols · 2024Review
- CRISETR: an efficient technology for multiplexed refactoring of biosynthetic gene clusters.Nucleic acids research · 2024Article
- Biosynthesis and engineering of the nonribosomal peptides with a C-terminal putrescine.Nature communications · 2023Article
- High-efficiency genetic engineering toolkit for virus based on lambda red-mediated recombination.Biotechnology letters · 2023Article
- Recent advances in the direct cloning of large natural product biosynthetic gene clusters.Engineering microbiology · 2023Review
- The emerging role of recombineering in microbiology.Engineering microbiology · 2023Review
- ClusterCAD 2.0: an updated computational platform for chimeric type I polyketide synthase and nonribosomal peptide synthetase design.Nucleic acids research · 2023Article
- Direct cloning of a herpesvirus genome for rapid generation of infectious BAC clones.Journal of advanced research · 2023Article
- Review
- Engineering ofFrontiers in microbiology · 2022Article
- Bacterioferritin: a key iron storage modulator that affects strain growth and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona.Microbial cell factories · 2021Article
- Recent Advances in Silent Gene Cluster Activation inFrontiers in bioengineering and biotechnology · 2021Review
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Authors and funding
14 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Biosynthesis reprograming is an important way to diversify chemical structures. The large repetitive DNA sequences existing in polyketide synthase genes make seamless DNA manipulation of the polyketide biosynthetic gene clusters extremely challenging. In this study, to replace the ethyl group attached to the C-21 of the macrolide insecticide spinosad with a butenyl group by refactoring the 79-kb gene cluster, we developed a RedEx method by combining Redαβ mediated linear-circular homologous recombination, ccdB counterselection and exonuclease mediated in vitro annealing to insert an exogenous extension module in the polyketide synthase gene without any extra sequence. RedEx was also applied for seamless deletion of the rhamnose 3'-O-methyltransferase gene in the spinosad gene cluster to produce rhamnosyl-3'-desmethyl derivatives. The advantages of RedEx in seamless mutagenesis will facilitate rational design of complex DNA sequences for diverse purposes.
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