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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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A reversible small-molecule-switchable self-amplifying RNA expression platform.

Guibin Lin1, Runjun Chen1, Zhengqiang Fu1

  • 1School of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou, Guangdong, 511442, China; National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, Guangzhou, Guangdong 510006, China; Guangdong Provincial Key Laboratory of Biomedical Engineering, South China University of Technology, Guangzhou, Guangdong, 510006, China; Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou, Guangdong, 510006, China.

International Journal of Biological Macromolecules
|March 16, 2026
PubMed
Summary
This summary is machine-generated.

This study engineered self-amplifying RNA (saRNA) as a programmable gene circuit. Small molecules like doxycycline and trimethoprim can now precisely control gene expression on and off, advancing RNA therapeutics.

Keywords:
Programmable RNA gene circuitsSelf-amplifying RNASmall-molecule-switchable gene expression

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • RNA Therapeutics

Background:

  • Precise control of gene expression is crucial for RNA therapeutics.
  • Self-amplifying RNA (saRNA) offers potential for modular gene regulation.
  • saRNA's capability as a programmable gene circuit for small-molecule control is underexplored.

Purpose of the Study:

  • To engineer saRNA constructs for small-molecule-controlled, multi-gene expression.
  • To establish saRNA as a programmable RNA gene circuit platform.
  • To evaluate the in vitro and in vivo efficacy of ligand-responsive saRNA.

Main Methods:

  • Engineered saRNA with regulatory modules (DD, L7Ae, TetR, k-turn).
  • Incorporated multiple subgenomic promoters for independent gene regulation.
  • Encapsulated saRNA into lipid nanoparticles for delivery and evaluated reporter gene expression.

Main Results:

  • Demonstrated independent and reversible gene regulation by trimethoprim (TMP) and doxycycline (Dox).
  • Showcased TMP and Dox as molecular switches for autonomous gene expression control.
  • Validated saRNA-mediated expression of reporter proteins in vitro and in vivo.

Conclusions:

  • Established saRNA as a programmable gene circuit with ligand-responsive, multi-gene regulatory capacity.
  • Provides a foundation for developing controllable RNA therapeutics.
  • Highlights potential for diverse synthetic biology applications.