ArticleNucleic acids research2014
Simultaneous non-contiguous deletions using large synthetic DNA and site-specific recombinases.
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 12 papers.
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Who cites it
12 citing papers in PubMed, 27 citations in OpenAlex.
- Robust and highly efficient transformation method for a minimal mycoplasma cell.Journal of bacteriology · 2025Article
- Review
- Creating custom synthetic genomes in Escherichia coli with REXER and GENESIS.Nature protocols · 2021Article
- Reprogramming the genetic code.Nature reviews. Genetics · 2021Review
- Article
- Coupling the recombineering to Cre-lox system enables simplified large-scale genome deletion in Lactobacillus casei.Microbial cell factories · 2018Article
- The Blueprint of a Minimal Cell: MiniBacillus.Microbiology and molecular biology reviews : MMBR · 2016Review
- Article
- Targeted Large-Scale Deletion of Bacterial Genomes Using CRISPR-Nickases.ACS synthetic biology · 2015Article
- Bacterial genome reduction using the progressive clustering of deletions via yeast sexual cycling.Genome research · 2015Article
- Strategies for cloning and manipulating natural and synthetic chromosomes.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2015Review
- Recombinase-mediated cassette exchange (RMCE) system for functional genomics studies in Mycoplasma mycoides.Biological procedures online · 2015Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Toward achieving rapid and large scale genome modification directly in a target organism, we have developed a new genome engineering strategy that uses a combination of bioinformatics aided design, large synthetic DNA and site-specific recombinases. Using Cre recombinase we swapped a target 126-kb segment of the Escherichia coli genome with a 72-kb synthetic DNA cassette, thereby effectively eliminating over 54 kb of genomic DNA from three non-contiguous regions in a single recombination event. We observed complete replacement of the native sequence with the modified synthetic sequence through the action of the Cre recombinase and no competition from homologous recombination. Because of the versatility and high-efficiency of the Cre-lox system, this method can be used in any organism where this system is functional as well as adapted to use with other highly precise genome engineering systems. Compared to present-day iterative approaches in genome engineering, we anticipate this method will greatly speed up the creation of reduced, modularized and optimized genomes through the integration of deletion analyses data, transcriptomics, synthetic biology and site-specific recombination.
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Registered trials
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