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DNA Isolation and Restriction Enzyme Analysis
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Restriction Enzymes as a Target for DNA-Based Sensing and Structural Rearrangement
Susan Buckhout-White1, Chanel Person1, Igor L Medintz1
1Center for Bio/Molecular Science and Engineering, Code 6900, U.S. Naval Research Laboratory, Washington, DC 20375, United States.
ACS Omega
|August 29, 2019
Summary
This study integrates restriction enzymes with DNA nanostructures for advanced sensing. The novel DNA switch detects multiple enzyme inputs, enabling complex logic-enabled sensing applications.
Area of Science:
- Biotechnology
- Nanotechnology
- Molecular Biology
Background:
- DNA nanostructures offer potential for complex logic-enabled sensing motifs.
- Current DNA sensing devices are primarily limited to DNA-based inputs.
- Restriction endonucleases possess inherent DNA specificity, making them suitable enzyme targets for triggering DNA structural changes.
Purpose of the Study:
- To investigate the integration of restriction endonucleases as inputs for DNA nanostructure-based sensing.
- To reconfigure a three-arm DNA switch to detect restriction endonuclease activity.
- To explore the potential for enzyme-triggered structural rearrangement in DNA nanostructures.
Main Methods:
- Reconfiguration of a three-arm DNA switch utilizing cyclic Förster resonance energy transfer (FRET) between three dyes.
- Design of specific DNA cleavage sites for three distinct restriction endonucleases.
- Investigation of enzyme target efficacy and specificity within the DNA switch architecture.
Main Results:
- Demonstrated successful detection of three different restriction enzymes using a single DNA switch.
- Confirmed no non-specific interactions between the cleavage sites of the three enzymes.
- Showcased enzymatic digestion's ability to expose an active toehold for subsequent oligo addition.
Conclusions:
- Restriction endonucleases can be effectively utilized as inputs for logic-enabled DNA nanostructure sensing.
- The developed DNA switch provides a versatile platform for multiplexed enzymatic detection.
- Enzyme-mediated structural changes in DNA nanostructures can be harnessed for programmable molecular logic and sensing.
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