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

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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

Fingerprint

Yeast
Genes
Yeasts
Genome
Oligonucleotides
Genetic Recombination
Gene Knockout Techniques
Genomic Library
Endonucleases
Eukaryota
Gene Frequency
Saccharomyces cerevisiae
Alleles
Screening
Population

Keywords

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

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
Dicarlo, James E. ; Conley, Andrew J. ; Penttilä, Merja ; Jäntti, Jussi ; Wang, Harris H. ; Church, George M. / Yeast oligo-mediated genome engineering (YOGE). In: ACS Synthetic Biology. 2013 ; Vol. 2, No. 12. pp. 741-749.
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Dicarlo, JE, Conley, AJ, Penttilä, M, Jäntti, J, Wang, HH & Church, GM 2013, 'Yeast oligo-mediated genome engineering (YOGE)', ACS Synthetic Biology, vol. 2, no. 12, pp. 741-749. https://doi.org/10.1021/sb400117c

Yeast oligo-mediated genome engineering (YOGE). / Dicarlo, James E.; Conley, Andrew J.; Penttilä, Merja; Jäntti, Jussi; Wang, Harris H.; Church, George M. (Corresponding Author).

In: ACS Synthetic Biology, Vol. 2, No. 12, 20.12.2013, p. 741-749.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Church, George M.

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