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Creating functional chromosome fusions in yeast with CRISPR-Cas9
Yangyang Shao1,2, Ning Lu1,2, Xiaoli Xue3
1Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, China.
Nature Protocols
|July 14, 2019
Summary
Researchers created a single-chromosome yeast strain using CRISPR-Cas9 chromosome fusion. This new method simplifies studying chromosome biology, including replication and segregation, in a functional, reduced genome.
Area of Science:
- Molecular Biology
- Genetics
- Yeast Biology
Background:
- Studying fundamental chromosome biology requires simplified genetic systems.
- Existing methods for chromosome manipulation are complex and time-consuming.
- CRISPR-Cas9 technology offers precise genome editing capabilities.
Purpose of the Study:
- To develop a protocol for CRISPR-Cas9-facilitated functional chromosome fusion in yeast.
- To generate a yeast strain with a single chromosome for biological studies.
- To provide guidelines for accurate and functional chromosome fusions.
Main Methods:
- Deletion of long redundant repetitive sequences near chromosomal ends.
- Determination of precise fusion order and cleavage sites.
- Minimization of gene expression influence to maintain functionality.
Main Results:
- Successfully created a yeast strain with a single functional chromosome.
- Established a protocol for generating functional chromosome fusions.
- Demonstrated that chromosome fusion can be achieved within 18 days.
Conclusions:
- CRISPR-Cas9 chromosome fusion is a viable method for generating reduced chromosome number yeast strains.
- This protocol facilitates the study of essential chromosome biology processes.
- The generated single-chromosome yeast serves as a valuable resource for genetic research.
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