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CRI-SPA: a high-throughput method for systematic genetic editing of yeast libraries
Paul Cachera1, Helén Olsson1, Hilde Coumou2
1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Denmark.
Nucleic Acids Research
|August 12, 2023
Summary
We developed CRI-SPA, a rapid method for mapping genetic features to function in yeast. This tool efficiently transfers genes, enabling genome-wide studies of biological processes like betaxanthin production.
Area of Science:
- Systems Biology
- Synthetic Biology
- Genetics
Background:
- Biological functions rely on complex genetic interaction networks.
- Predicting these interactions is crucial for systems biology but remains challenging.
- Novel tools for rapid sequence-to-function mapping are needed.
Purpose of the Study:
- To introduce CRI-SPA, a novel method for high-throughput genetic feature transfer in Saccharomyces cerevisiae.
- To demonstrate the efficiency and reproducibility of CRI-SPA for large-scale functional genomics.
Main Methods:
- CRI-SPA combines mating, CRISPR-Cas9-induced gene conversion, and Selective Ploidy Ablation.
- The method allows massively parallelized, automated transfer of chromosomal genetic features.
- Applied to transfer betaxanthin production genes into the yeast knockout collection (≈4800 strains).
Main Results:
- CRI-SPA demonstrated high efficiency and reproducibility in transferring genetic features.
- The method successfully enabled marker-free transfer of genes.
- Validated hits showed strong correlation between transferred features and recreated mutant phenotypes, providing a genome-wide view of betaxanthin production requirements.
Conclusions:
- CRI-SPA is a versatile, rapid, and reliable tool for systems-level understanding of biological processes.
- Facilitates forward genetics and functional genomics in yeast.
- Enables efficient genome-wide screening for genetic requirements of complex traits.
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