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A generalizable approach for programming protease-responsive conformationally inhibited artificial transcriptional
Yinxia Liu1, Lingyun Zhao1, Jinshan Long1
1Affiliated XiangTan Central Hospital of Hunan University, College of Chemistry and Chemical Engineering, School of Biomedical Sciences, Hunan University, Changsha, 410082, China.
Nature Communications
|May 17, 2025
Summary
Researchers developed a protease-responsive system (PRCIS) to control synthetic genetic circuits. This system uses artificial transcription factors (ATFs) that are activated by specific protease cleavage, enabling precise control over gene expression for synthetic biology applications.
Area of Science:
- Synthetic biology
- Molecular and cell biology
- Biochemistry
Background:
- Synthetic genetic circuits are crucial for cell biology research.
- Artificial transcription factors (ATFs) are key components for building programmable gene expression systems.
- Controlling ATF activity is essential for orthogonal and composable transcriptional regulation.
Purpose of the Study:
- To design a novel protease-responsive conformationally inhibited system (PRCIS).
- To enable event-triggerable cascaded pathways using ATFs.
- To provide a generalizable design for advanced synthetic biology applications.
Main Methods:
- Intramolecularly linking ATF DNA-binding domains to confined dimerized regions to create conformational inhibition.
- Utilizing proteolytic cleavage to release inhibition and restore DNA binding.
- Adapting the PRCIS design to various ATFs and proteases.
Main Results:
- Demonstrated successful conformational inactivation and reactivation of ATF DNA binding.
- Achieved high activation ratios and specificity in protease-responsive gene expression.
- Developed PRCIS-based platforms for triple-orthogonal protease-responsive and dual-orthogonal chemical-inducible systems.
- Implemented Boolean logic operations using the PRCIS platform.
Conclusions:
- PRCIS offers a versatile and generalizable design for controlling synthetic genetic circuits.
- The system enables precise, event-triggered gene expression through protease-mediated activation.
- PRCIS advances the development of sophisticated synthetic biology tools for research and applications.
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