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Updated: Sep 25, 2025
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Automated Robotic Liquid Handling Assembly of Modular DNA Devices
Published on: December 1, 2017
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Engineering protein activity into off-the-shelf DNA devices.
Harsimranjit Sekhon1, Stewart N Loh1
1Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, Syracuse, NY 13210, USA.
Cell Reports Methods
|May 2, 2022
Summary
Researchers engineered a novel protein switch (nLuc-AFF) activated by DNA. This DNA-activated enzyme enables versatile biosensors and logic gates, even in crude biological samples.
Area of Science:
- Biotechnology
- Molecular Engineering
- Synthetic Biology
Background:
- DNA devices offer predictable design but lack biological functions like catalysis.
- Protein devices provide diverse functions but are challenging to engineer due to complex folding.
- Integrating DNA and protein engineering can overcome these limitations.
Purpose of the Study:
- To engineer a DNA-activated enzyme switch for biosensing and logic gate applications.
- To develop a versatile platform combining DNA programmability with protein functionality.
- To create a genetically encoded, ratiometric biosensor suitable for crude samples.
Main Methods:
- Engineered a protein switch (nLuc-AFF) from nanoluciferase using the alternate-frame-folding mechanism.
- Paired nLuc-AFF with DNA technologies to create biosensors and logic gates.
- Utilized blue/green luminescence for ratiometric readout quantifiable by phone camera.
Main Results:
- Developed nLuc-AFF, a DNA-sequence-activated protein switch.
- Demonstrated nLuc-AFF's utility in creating nucleic acid sensors, serotonin/ATP sensors, and a two-input logic gate.
- Confirmed nLuc-AFF's ratiometric readout stability in 100% serum for crude sample analysis.
Conclusions:
- The nLuc-AFF system provides a robust method for creating DNA-activated biosensors and logic gates.
- This approach allows for the conversion of various proteins and enzymes into DNA-activated switches.
- The technology is suitable for low-resource settings, enabling versatile biological sensing and computation.
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