Yeast oligo-mediated genome engineering (YOGE)

James E. Dicarlo, Andrew J. Conley, Merja Penttilä, Jussi Jäntti, Harris H. Wang, George M. Church (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

93 Citations (Scopus)

Abstract

High-frequency oligonucleotide-directed recombination engineering (recombineering) has enabled rapid modification of several prokaryotic genomes to date. Here, we present a method for oligonucleotide-mediated recombineering in the model eukaryote and industrial production host Saccharomyces cerevisiae, which we call yeast oligo-mediated genome engineering (YOGE). Through a combination of overexpression and knockouts of relevant genes and optimization of transformation and oligonucleotide designs, we achieve high gene-modification frequencies at levels that only require screening of dozens of cells. We demonstrate the robustness of our approach in three divergent yeast strains, including those involved in industrial production of biobased chemicals. Furthermore, YOGE can be iteratively executed via cycling to generate genomic libraries up to 105 individuals at each round for diversity generation. YOGE cycling alone or in combination with phenotypic selections or endonuclease-based negative genotypic selections can be used to generate modified alleles easily in yeast populations with high frequencies.

Original languageEnglish
Pages (from-to)741-749
JournalACS Synthetic Biology
Volume2
Issue number12
DOIs
Publication statusPublished - 20 Dec 2013
MoE publication typeA1 Journal article-refereed

Keywords

  • genome engineering
  • MAGE
  • oligonucleotide transformation
  • recombineering
  • Saccharomyces cerevisiae
  • yeast

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  • Cite this

    Dicarlo, J. E., Conley, A. J., Penttilä, M., Jäntti, J., Wang, H. H., & Church, G. M. (2013). Yeast oligo-mediated genome engineering (YOGE). ACS Synthetic Biology, 2(12), 741-749. https://doi.org/10.1021/sb400117c