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Efficient site-specific recombination by self-activating split-Dre recombinase in mammalian cells and E. coli
Chichu Xie1,2, Jinfeng Gan3, Yuanhao Zhang3
1Guangxi Key Laboratory of Tumor Immunology and Microenvironmental Regulation, Guilin Medical University, Guilin, 541199, China. xcc335@glmu.edu.cn.
Journal of Biological Engineering
|September 2, 2025
Summary
Researchers developed a self-activating split Dre recombinase (N191/192C) for efficient site-specific recombination. This novel system functions without external signals and is compatible with split Cre, offering new gene editing tools.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- Site-specific recombination (SSR) systems are crucial for genetic manipulation.
- Novel SSR strategies are needed for advanced applications.
- This study focused on developing a self-activating split Dre recombinase.
Purpose of the Study:
- To identify a self-activating split Dre protein configuration.
- To evaluate the recombinase activity of split Dre variants.
- To assess the compatibility of split Dre with split Cre systems.
Main Methods:
- Designed and tested various split Dre protein fragments.
- Assessed recombination efficiency between rox sites in different cell lines and bacteria.
- Investigated the effect of intein fusion on split Dre activity.
Main Results:
- The N191/192C split Dre pair showed significant self-activating recombinase activity.
- This combination efficiently mediated site-specific recombination at integrated rox sites.
- No cross-complementation was observed between split Dre and split Cre proteins.
Conclusions:
- The N191/192C split Dre protein offers a self-activating system for recombination.
- Split Dre and Cre systems can be used concurrently without interference.
- These findings provide novel tools for gene editing and transgenic animal generation.
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