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TALENs-Assisted Multiplex Editing for Accelerated Genome Evolution To Improve Yeast Phenotypes.
Guoqiang Zhang1,2, Yuping Lin1, Xianni Qi1
1Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences , Tianjin 300308, China.
ACS Synthetic Biology
|May 27, 2015
Summary
Transcription activator-like effector nucleases (TALENs)-assisted multiplex editing (TAME) enables rapid, large-scale genome modifications. This platform accelerates yeast genome evolution for improved traits like ethanol tolerance.
Area of Science:
- Molecular Biology
- Synthetic Biology
- Genetics
Background:
- Genome editing is crucial for creating desired genotypes and phenotypes.
- A key challenge is achieving rapid, genome-wide genetic alterations.
- Transcription activator-like effector nucleases (TALENs) offer targeted DNA modification capabilities.
Purpose of the Study:
- To introduce TALENs-assisted multiplex editing (TAME) for efficient, large-scale genome modifications.
- To demonstrate TAME's utility in rapidly improving yeast phenotypes through iterative cycles.
- To evaluate TAME's effectiveness in enhancing ethanol tolerance in yeast.
Main Methods:
- Designed TALENs targeting DNA sequences between TATA and GC boxes.
- Iterative TAME cycles to induce targeted genomic modifications (indels).
- Reporter-assisted screening for efficient selection of improved yeast strains.
Main Results:
- TAME successfully generated multiple targeted genomic modifications.
- Iterative TAME and screening rapidly improved a fluorescent trait in yeast.
- Significant enhancement of yeast ethanol tolerance was achieved in a short period.
Conclusions:
- TAME is a broadly generalizable platform for accelerated genome evolution.
- The TAME system enables rapid improvement of yeast phenotypes.
- This method facilitates the development of yeast strains with desirable industrial traits.
