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Updated: May 31, 2026
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DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
Published on: December 29, 2021
A T7 RNAP regulatory toolbox for cell-free network engineering and biosensing applications
Pao-Wan Lee1, Seyed Saeed Mottaghi1, Matthis Guillaume Lugnier1
1Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Nature Communications
|May 28, 2026
Summary
We developed a versatile synthetic biology toolbox to precisely control T7 RNA polymerase (RNAP) for gene circuits. This system enables sensitive detection of various biomolecules and potential point-of-care diagnostics.
Area of Science:
- Synthetic biology
- Biotechnology
- Molecular engineering
Background:
- T7 RNA polymerase (RNAP) is widely used but lacks precise regulatory control.
- Limited modularity hinders the design of complex T7 RNAP-based gene regulatory networks.
Purpose of the Study:
- To engineer a programmable toolbox for T7 RNAP regulation in cell-free systems.
- To enable scalable design of gene circuits and sensitive biomolecule detection.
- To develop rapid, point-of-care diagnostic assays.
Main Methods:
- Engineered synthetic repressors, activators, and biosensors for T7 RNAP.
- Integrated a protein design pipeline for synthetic binder generation.
- Developed a cell-free system for multiplexed detection and diagnostics.
Main Results:
- Demonstrated scalable design of T7 RNAP-based gene regulatory networks.
- Achieved rapid, sensitive, and multiplexed detection of small molecules, antibodies, and proteins.
- Constructed a SARS-CoV-2 diagnostic assay with high sensitivity using combined amplification strategies.
Conclusions:
- The developed toolbox provides a flexible and expandable framework for engineering biomolecule-responsive gene circuits.
- The cell-free system shows significant potential for developing point-of-care diagnostic assays.
