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Updated: May 13, 2026
08:57
High-throughput Yeast Plasmid Overexpression Screen
Published on: July 27, 2011
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Modular RNAi Pathway Engineering Enhances Plasmid Copy Number Control in Yeast Bioproduction System
Qianru Cai1, Manman Wang1, Jinmei Zhu1
1Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou, Zhejiang, China.
Biotechnology and Bioengineering
|June 30, 2025
Summary
Scientists engineered a dynamic plasmid copy number system in yeast using RNA interference (RNAi). This synthetic biology tool boosts gene dosage control and enhances microbial cell factory production.
Area of Science:
- Synthetic Biology
- Metabolic Engineering
- Molecular Biology
Background:
- Optimizing metabolic flux in microbial cell factories requires precise control over gene dosage and expression dynamics.
- Cellular burden must be minimized to ensure efficient production.
- Dynamic regulation of gene expression is crucial for advanced metabolic engineering.
Purpose of the Study:
- To develop a synthetic biology chassis in Saccharomyces cerevisiae with dynamically programmable plasmid copy numbers.
- To establish a chemically inducible platform for precise gene dosage control.
- To enhance the production of valuable compounds in microbial cell factories.
Main Methods:
- Integrated heterologous RNA interference (RNAi) pathway genes from Saccharomyces castellii.
- Designed sequence-specific small interfering RNAs (siRNAs) targeting plasmid-encoded selection markers.
- Developed a chemically inducible system for regulating plasmid copy number.
Main Results:
- Achieved up to a 7.13-fold amplification in plasmid copy number.
- Demonstrated an 18.6-fold increase in lycopene titers by modulating carotenoid biosynthesis.
- Established a novel RNAi-mediated gene dosage control platform.
Conclusions:
- The developed RNAi-mediated system enables dynamic plasmid copy number regulation in S. cerevisiae.
- This strategy significantly enhances production performance in microbial cell factories.
- Offers new perspectives for metabolic engineering and synthetic biology applications.
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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
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This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
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