ArticleFEMS yeast research2013
One-step assembly and targeted integration of multigene constructs assisted by the I-SceI meganuclease in Saccharomyces cerevisiae.
Article in FEMS yeast research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Functional dissection ofProceedings of the National Academy of Sciences of the United States of America · 2026Article
- Site-Directed Genome Integration via Recombinase-Mediated Cassette Exchange (RMCE) inACS synthetic biology · 2025Article
- Recent technology advancements in large-scale DNA assembly.Quantitative biology (Beijing, China) · 2025Article
- The Construction of Heterothallic Strains ofBiomolecules · 2025Article
- Cyanamide-inducible expression of homing nucleaseSynthetic and systems biotechnology · 2024Article
- Acceleration of CRISPR/Cas9-Mediated Editing at Multiple Sites in theMethods and protocols · 2023Article
- A Versatile in Vivo DNA Assembly Toolbox for Fungal Strain Engineering.ACS synthetic biology · 2022Article
- Construction of a synthetic Saccharomyces cerevisiae pan-genome neo-chromosome.Nature communications · 2022Article
- CRISPR-Cas9: A Powerful Tool to Efficiently EngineerLife (Basel, Switzerland) · 2020Review
- Laboratory evolution of a glucose-phosphorylation-deficient, arabinose-fermenting S. cerevisiae strain reveals mutations in GAL2 that enable glucose-insensitive l-arabinose uptake.FEMS yeast research · 2018Article
- Genome editing in Kluyveromyces and Ogataea yeasts using a broad-host-range Cas9/gRNA co-expression plasmid.FEMS yeast research · 2018Article
- FnCpf1: a novel and efficient genome editing tool for Saccharomyces cerevisiae.Nucleic acids research · 2017Article
- Integrative modules for efficient genome engineering in yeast.Microbial cell (Graz, Austria) · 2017Article
- Mutations in PMR1 stimulate xylose isomerase activity and anaerobic growth on xylose of engineered Saccharomyces cerevisiae by influencing manganese homeostasis.Scientific reports · 2017Article
- CASCADE, a platform for controlled gene amplification for high, tunable and selection-free gene expression in yeast.Scientific reports · 2017Article
- Pathway swapping: Toward modular engineering of essential cellular processes.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Technology development for natural product biosynthesis in Saccharomyces cerevisiae.Current opinion in biotechnology · 2016Review
- EasyClone-MarkerFree: A vector toolkit for marker-less integration of genes into Saccharomyces cerevisiae via CRISPR-Cas9.Biotechnology journal · 2016Article
- Combinatorial metabolic pathway assembly in the yeast genome with RNA-guided Cas9.Journal of industrial microbiology & biotechnology · 2016Article
- I-SceI-mediated double-strand DNA breaks stimulate efficient gene targeting in the industrial fungus Trichoderma reesei.Applied microbiology and biotechnology · 2015Article
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8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In vivo assembly of overlapping fragments by homologous recombination in Saccharomyces cerevisiae is a powerful method to engineer large DNA constructs. Whereas most in vivo assembly methods reported to date result in circular vectors, stable integrated constructs are often preferred for metabolic engineering as they are required for large-scale industrial application. The present study explores the potential of combining in vivo assembly of large, multigene expression constructs with their targeted chromosomal integration in S. cerevisiae. Combined assembly and targeted integration of a ten-fragment 22-kb construct to a single chromosomal locus was successfully achieved in a single transformation process, but with low efficiency (5% of the analyzed transformants contained the correctly assembled construct). The meganuclease I-SceI was therefore used to introduce a double-strand break at the targeted chromosomal locus, thus to facilitate integration of the assembled construct. I-SceI-assisted integration dramatically increased the efficiency of assembly and integration of the same construct to 95%. This study paves the way for the fast, efficient, and stable integration of large DNA constructs in S. cerevisiae chromosomes.
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