ArticleNucleic acids research2010
Tandem repeat coupled with endonuclease cleavage (TREC): a seamless modification tool for genome engineering in yeast.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 50 citations in OpenAlex.
- Article
- An Expanded Genetic Toolbox to Accelerate the Creation ofACS synthetic biology · 2024Article
- Cloning and sequencing analysis of wholeFrontiers in microbiology · 2024Article
- SLICER: A Seamless Gene Deletion Method forBiodesign research · 2023Article
- Gene editing tools for mycoplasmas: references and future directions for efficient genome manipulation.Frontiers in microbiology · 2023Review
- Genome Engineering of the Fast-GrowingACS synthetic biology · 2022Article
- Debugging: putting the synthetic yeast chromosome to work.Chemical science · 2021Review
- Budding yeast as a factory to engineer partial and complete microbial genomes.Current opinion in systems biology · 2020Review
- Article
- Evidence for the Cytoplasmic Localization of the L-α-Glycerophosphate Oxidase in Members of the "Frontiers in microbiology · 2019Article
- Removal of a Subset of Non-essential Genes Fully Attenuates a Highly VirulentFrontiers in microbiology · 2019Article
- Impact of donor-recipient phylogenetic distance on bacterial genome transplantation.Nucleic acids research · 2016Article
- 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
- TREC-IN: gene knock-in genetic tool for genomes cloned in yeast.BMC genomics · 2014Article
- Simultaneous non-contiguous deletions using large synthetic DNA and site-specific recombinases.Nucleic acids research · 2014Article
- The flavoprotein Mcap0476 (RlmFO) catalyzes m5U1939 modification in Mycoplasma capricolum 23S rRNA.Nucleic acids research · 2014Article
- Transferring whole genomes from bacteria to yeast spheroplasts using entire bacterial cells to reduce DNA shearing.Nature protocols · 2014Article
- Assembly of eukaryotic algal chromosomes in yeast.Journal of biological engineering · 2013Article
- One-step assembly and targeted integration of multigene constructs assisted by the I-SceI meganuclease in Saccharomyces cerevisiae.FEMS yeast research · 2013Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The complete synthetic Mycoplasma genitalium genome ( approximately 583 kb) has been assembled and cloned as a circular plasmid in the yeast Saccharomyces cerevisiae. Attempts to engineer the cloned genome by standard genetic methods involving the URA3/5-fluoroorotic acid (5-FOA) counter-selection have shown a high background of 5-FOA resistant clones derived from spontaneous deletions of the bacterial genome maintained in yeast. Here, we report a method that can seamlessly modify the bacterial genome in yeast with high efficiency. This method requires two sequential homologous recombination events. First, the target region is replaced with a mutagenesis cassette that consists of a knock-out CORE (an18-bp I-SceI recognition site, the SCEI gene under the control of the GAL1 promoter, and the URA3 marker) and a DNA fragment homologous to the sequence upstream of the target site. The replacement generates tandem repeat sequences flanking the CORE. Second, galactose induces the expression of I-SceI, which generates a double-strand break (DSB) at the recognition site. This DSB promotes intra-molecular homologous recombination between the repeat sequences, and leads to an excision of the CORE. As a result, a seamless modification is generated. This method can be adapted for a variety of genomic modifications and may provide an important tool to modify and design natural or synthetic genomes propagated in yeast.
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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.