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Engineering Cell-permeable Protein
Published on: December 28, 2009
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A chemically inducible, leakless Cre recombinase by split-protein-based efficient and enhanced degradation (SPEED)
Dewen Cai1, Fuun Kawano1,2, Takahiro Otabe1,3
1Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo, Japan.
Communications Biology
|December 14, 2025
Summary
Researchers developed a new system called SPEED-Cre for precise genetic control. This leakless system uses a destabilized Cre recombinase activated by trimethoprim (TMP), enabling efficient DNA recombination in cells and living organisms.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- Chemically inducible DNA recombination systems are crucial for genetic applications.
- Conventional systems exhibit undesirable leaky activity without chemical induction.
Purpose of the Study:
- To develop a novel, leakless chemically inducible DNA recombination system.
- To engineer a destabilized Cre recombinase (SPEED-Cre) with enhanced control.
Main Methods:
- Utilized a split-protein-based efficient and enhanced degradation (SPEED) approach.
- Fused a destabilizing domain mutant of E. coli dihydrofolate reductase to split-Cre.
- Stabilized the construct using the antibiotic trimethoprim (TMP).
Main Results:
- Demonstrated no significant background DNA recombination in the absence of TMP.
- Achieved full TMP-dependent Cre-loxP recombination in human cells and mice.
- Showcased the SPEED approach's applicability to other recombinases (Flp, VCre, Dre).
Conclusions:
- SPEED-Cre offers a robust, leakless platform for precise chemogenetic control.
- This technology significantly advances genome engineering in living systems.
- The SPEED platform has broad potential for various protein applications.
