ArticleNucleic acids research2017
Large-scale recoding of a bacterial genome by iterative recombineering of synthetic DNA.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Who cites it
46 citing papers in PubMed.
- Firewalled synthetic commensal blocks horizontal gene transfer in the gut.bioRxiv : the preprint server for biology · 2026Article
- Probing the limits of genetic recoding using multi-omics-guided evolution.Nature communications · 2026Article
- Automated linear DNA assembly of A. thaliana's chloroplast and mitochondrial genome.Nucleic acids research · 2026Article
- Advances in large-scale DNA engineering with the CRISPR system.Experimental & molecular medicine · 2025Review
- The design and engineering of synthetic genomes.Nature reviews. Genetics · 2025Review
- XanthoMoClo─A Robust Modular Cloning Genetic Toolkit for the GeneraACS synthetic biology · 2025Article
- Article
- Genomically recodedbioRxiv : the preprint server for biology · 2024Article
- Synthetic genomes unveil the effects of synonymous recoding.bioRxiv : the preprint server for biology · 2024Article
- Genome engineering on size reduction and complexity simplification: A review.Journal of advanced research · 2024Review
- Microbial Catalysis for COCold Spring Harbor perspectives in biology · 2024Review
- Robust genetic codes enhance protein evolvability.PLoS biology · 2024Article
- Engineering bacteria for cancer immunotherapy.Current opinion in biotechnology · 2024Review
- Strategies to identify and edit improvements in synthetic genome segments episomally.Nucleic acids research · 2023Article
- Enabling technology and core theory of synthetic biology.Science China. Life sciences · 2023Review
- Article
- Multiplex base editing to convert TAG into TAA codons in the human genome.Nature communications · 2022Article
- Current trends in biopharmaceuticals production in Escherichia coli.Biotechnology letters · 2022Review
- Review
- Quantification of synthetic errors during chemical synthesis of DNA and its suppression by non-canonical nucleosides.Scientific reports · 2022Article
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Authors and funding
13 authors.
Funding
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Abstract
The ability to rewrite large stretches of genomic DNA enables the creation of new organisms with customized functions. However, few methods currently exist for accumulating such widespread genomic changes in a single organism. In this study, we demonstrate a rapid approach for rewriting bacterial genomes with modified synthetic DNA. We recode 200 kb of the Salmonella typhimurium LT2 genome through a process we term SIRCAS (stepwise integration of rolling circle amplified segments), towards constructing an attenuated and genetically isolated bacterial chassis. The SIRCAS process involves direct iterative recombineering of 10-25 kb synthetic DNA constructs which are assembled in yeast and amplified by rolling circle amplification. Using SIRCAS, we create a Salmonella with 1557 synonymous leucine codon replacements across 176 genes, the largest number of cumulative recoding changes in a single bacterial strain to date. We demonstrate reproducibility over sixteen two-day cycles of integration and parallelization for hierarchical construction of a synthetic genome by conjugation. The resulting recoded strain grows at a similar rate to the wild-type strain and does not exhibit any major growth defects. This work is the first instance of synthetic bacterial recoding beyond the Escherichia coli genome, and reveals that Salmonella is remarkably amenable to genome-scale modification.
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