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Yeast oligo-mediated genome engineering (YOGE).

James E DiCarlo1, Andrew J Conley, Merja Penttilä

  • 1Department of Genetics, Harvard Medical School , Boston, Massachusetts 02115, United States.

ACS Synthetic Biology
|October 29, 2013
PubMed
Summary

We developed yeast oligo-mediated genome engineering (YOGE), a new method for fast and efficient genetic modification in Saccharomyces cerevisiae. YOGE allows high-frequency gene editing in yeast strains, including those used for industrial bio-based chemical production.

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Yeast Genetics

Background:

  • Recombineering enables rapid genome modification in prokaryotes.
  • Efficient genome engineering in eukaryotes, particularly industrial yeast strains, is crucial for biotechnology.

Purpose of the Study:

  • To develop a high-frequency genome engineering method for Saccharomyces cerevisiae.
  • To adapt oligonucleotide-mediated recombineering for yeast, termed yeast oligo-mediated genome engineering (YOGE).

Main Methods:

  • Optimized gene knockouts, overexpression, transformation protocols, and oligonucleotide designs.
  • Applied YOGE to three divergent yeast strains, including industrial production hosts.
  • Developed iterative YOGE cycling for generating large genomic libraries.

Main Results:

  • Achieved high gene-modification frequencies in yeast, requiring screening of only dozens of cells.
  • Demonstrated YOGE's robustness across different yeast strains.
  • Generated genomic libraries of up to 10^5 individuals per round using YOGE cycling.

Conclusions:

  • YOGE provides a powerful and efficient tool for yeast genome engineering.
  • The method facilitates the generation of diverse modified yeast strains for bio-based chemical production.
  • Iterative YOGE cycling combined with selection strategies enables rapid creation of modified yeast populations.