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Updated: May 28, 2026
14:49
Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira
Published on: August 14, 2021
Tunable Transcription-Level CRISPR Interference in Vibrio natriegens Using Engineered Single Guide RNAs
Mincheol Choi1, Gibyuck Byun2, Giho Kim2
1Department of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul08826, Republic of Korea.
ACS Synthetic Biology
|May 26, 2026
Summary
Researchers developed a tunable CRISPR interference system for precise gene control in Vibrio natriegens. This advancement enhances its potential as a microbial platform for producing valuable chemicals.
Area of Science:
- Microbiology
- Synthetic Biology
- Metabolic Engineering
Background:
- Vibrio natriegens is a fast-growing bacterium with potential as a microbial production platform.
- Existing genetic tools for V. natriegens lack precise gene expression control.
- CRISPR interference (CRISPRi) offers a way to regulate gene expression.
Purpose of the Study:
- To develop a tunable CRISPR interference system for V. natriegens.
- To enable multilevel transcriptional regulation for enhanced biological engineering.
- To improve the production of value-added chemicals using V. natriegens.
Main Methods:
- Engineered single-guide RNA (sgRNA) variants by modifying tetraloop and flanking regions.
- Created a synthetic sgRNA library to modulate dCas9 binding affinity.
- Applied the tunable CRISPRi system for metabolic engineering in V. natriegens.
Main Results:
- Achieved modular gene repression across multiple targets using engineered sgRNAs.
- Demonstrated successful redirection of intracellular carbon flux.
- Increased 3-hydroxypropionic acid production by 2.2-fold and lycopene production by 1.5-fold.
Conclusions:
- Developed a simple and effective tunable CRISPRi system for V. natriegens.
- This system allows for precise, multilevel gene regulation.
- Expanded the utility of V. natriegens as a versatile platform for chemical production.
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