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Updated: Nov 15, 2025
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Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production
Published on: September 20, 2016
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Recombination machinery engineering for precise genome editing in methylotrophic yeast Ogataea polymorpha
Jiaoqi Gao1,2,3, Ning Gao1,3, Xiaoxin Zhai1,3
1Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, PR China.
Iscience
|March 5, 2021
Summary
Engineering recombination machinery in *Ogataea polymorpha* enhances homologous recombination (HR) efficiency. This breakthrough simplifies genetic engineering in yeasts, enabling efficient cell factory construction for industrial applications.
Area of Science:
- Synthetic biology
- Microbial biotechnology
Background:
- Methylotrophic yeasts like *Ogataea polymorpha* are valuable for biorefineries due to their ability to utilize methanol.
- Low homologous recombination (HR) efficiency in *O. polymorpha* limits precise genetic manipulation for cell factory development.
Purpose of the Study:
- To enhance the homologous recombination (HR) efficiency in *O. polymorpha* using recombination machinery engineering (rME).
- To establish an efficient CRISPR-Cas9 system for precise genetic engineering in this methylotrophic yeast.
Main Methods:
- Engineering the recombination machinery by overexpressing HR-related proteins and down-regulating non-homologous end joining (NHEJ).
- Implementing an efficient CRISPR-Cas9 system for targeted genetic modifications.
- Analyzing the competition between HR and NHEJ pathways in engineered mutants.
Main Results:
- Recombination machinery engineering significantly increased HR rates from 20%-30% to 60%-70%.
- Demonstrated homologous integration of large DNA fragments and *in vivo* assembly of multiple DNA fragments.
- Successfully engineered *O. polymorpha* for fatty alcohol production.
Conclusions:
- The developed HR-upregulated system simplifies genetic engineering in non-conventional yeasts.
- *O. polymorpha* is a promising and adaptable cell factory for industrial applications, facilitated by improved genetic tools.
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