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Ensemble Sensing Using Single-Molecule DNA Copolymers.

Sagun Jonchhe¹, Sangeetha Selvam¹, Deepak Karna¹

¹Department of Chemistry & Biochemistry and School of Biomedical Sciences, Kent State University, Kent, Ohio 44242, United States.

Analytical Chemistry

|August 25, 2020

Summary

This summary is machine-generated.

Researchers developed novel DNA copolymers for highly sensitive and rapid detection of microRNAs (miRNAs). This breakthrough combines single-molecule precision with ensemble assay accuracy, achieving femtomolar detection limits for miRNA analysis.

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Area of Science:

Biotechnology
Molecular Biology
Nanotechnology

Background:

Single-molecule sensing offers high mass sensitivity but suffers from slow detection times at low concentrations due to diffusion limitations.
Existing methods struggle with accuracy in determining concentration sensitivity for low-analyte levels.

Purpose of the Study:

To overcome the limitations of single-molecule sensing by developing a novel DNA-based material for accurate and sensitive analyte detection.
To achieve rapid, femtomolar detection limits for microRNAs (miRNAs).

Main Methods:

Utilized rolling circle amplification to create single-molecule DNA homopolymers with up to 473 identical DNA hairpin sensing elements.
Exploited ensemble unfolding/refolding transitions of DNA homopolymers for microRNA recognition.
Developed alternating DNA copolymers for multiplexed detection of different miRNAs.

Main Results:

DNA homopolymers with as few as 10 tandem hairpins exhibited ensemble transitions, enabling microRNA detection.
Achieved a femtomolar detection limit for miRNAs within 20 minutes, a 6-order magnitude improvement over standalone hairpins.
Demonstrated multiplex recognition of various miRNAs using alternating DNA copolymers.

Conclusions:

DNA homopolymers and copolymers represent innovative materials for advanced sensing strategies.
These materials successfully combine single-molecule precision with ensemble assay accuracy for concentration sensitivity determination.
The developed sensing platform offers a significant advancement in sensitive and rapid detection of miRNAs.