PMID:21764749
| Citation |
Isaacs, FJ, Carr, PA, Wang, HH, Lajoie, MJ, Sterling, B, Kraal, L, Tolonen, AC, Gianoulis, TA, Goodman, DB, Reppas, NB, Emig, CJ, Bang, D, Hwang, SJ, Jewett, MC, Jacobson, JM and Church, GM (2011) Precise manipulation of chromosomes in vivo enables genome-wide codon replacement. Science 333:348-53 |
|---|---|
| Abstract |
We present genome engineering technologies that are capable of fundamentally reengineering genomes from the nucleotide to the megabase scale. We used multiplex automated genome engineering (MAGE) to site-specifically replace all 314 TAG stop codons with synonymous TAA codons in parallel across 32 Escherichia coli strains. This approach allowed us to measure individual recombination frequencies, confirm viability for each modification, and identify associated phenotypes. We developed hierarchical conjugative assembly genome engineering (CAGE) to merge these sets of codon modifications into genomes with 80 precise changes, which demonstrate that these synonymous codon substitutions can be combined into higher-order strains without synthetic lethal effects. Our methods treat the chromosome as both an editable and an evolvable template, permitting the exploration of vast genetic landscapes. |
| Links |
PubMed Online version:10.1126/science.1205822 |
| Keywords |
Chromosomes, Bacterial/genetics; Codon, Terminator; Conjugation, Genetic; Directed Molecular Evolution; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli/physiology; Genetic Engineering/methods; Genome, Bacterial; Genomic Instability; Mutagenesis, Site-Directed; Mutation; Phenotype; Recombination, Genetic; Templates, Genetic |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
See also
References
See Help:References for how to manage references in EcoliWiki.