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Updated: Jun 10, 2026
08:35
Inducible and Reversible Dominant-negative (DN) Protein Inhibition
Published on: January 7, 2019
Selection systems based on dominant-negative transcription factors for precise genetic engineering
Raphaël Dutoit1, Evelyne Dubois, Eric Jacobs
1Institut de Recherches Microbiologiques JM Wiame and Laboratoire de Microbiologie de l'Université Libre de Bruxelles, 1 avenue Emile Gryson, BE1070 Belgium. rdutoit@ulb.ac.be
Nucleic Acids Research
|August 13, 2010
Summary
This study introduces a novel genetic engineering method for precise microbial genome modification. The approach uses dominant-negative transcription factors as selection markers, enabling targeted gene deletion without side effects.
Area of Science:
- Microbiology
- Molecular Biology
- Genetic Engineering
Background:
- Existing genetic modification tools lack precision, often causing unintended alterations.
- Precise genomic engineering is crucial for functional genomic analysis and industrial applications.
Purpose of the Study:
- To develop a new method for precise genomic engineering in microorganisms.
- To enable targeted gene modification without undesirable side-alterations.
Main Methods:
- Utilized truncated genes encoding dominant-negative transcription factors as selection markers.
- Demonstrated dominant-negative effects of truncated Gal4p and Arg81p in Saccharomyces cerevisiae.
- Applied these markers for precise gene deletion (HO and URA3) in wild yeasts.
Main Results:
- Successfully demonstrated dominant-negative effects of truncated transcription factors.
- Established a selection system using these markers for both positive and negative selection.
- Achieved precise deletion of target genes in yeast without off-target mutations.
Conclusions:
- The developed method allows for precise genomic engineering in microorganisms.
- The dominant-negative transcription factor system serves as an effective selection tool.
- This approach holds potential for broader application in microbial genetic engineering.
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