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Detection of miR-155 Using Peptide Nucleic Acid at Physiological-like Conditions by Surface Plasmon Resonance and
Francesco Lavecchia di Tocco1, Valentina Botti1, Salvatore Cannistraro1
1Biophysics and Nanoscience Centre, DEB, Università della Tuscia, Largo dell'Università, 01100 Viterbo, Italy.
Abstract:
MicroRNAs are small ribonucleotides that act as key gene regulators. Their altered expression is often associated with the onset and progression of several human diseases, including cancer. Given their potential use as biomarkers, there is a need to find detection methods for microRNAs suitable for use in clinical setting. Field-effect-transistor-based biosensors (bioFETs) appear to be valid tools to detect microRNAs, since they may reliably quantitate the specific binding between the immobilized probe and free target in solution through an easily detectable electrical signal. We have investigated the detection of human microRNA 155 (miR-155) using an innovative capturing probe constituted by a synthetic peptide nucleic acid (PNA), which has the advantage to form a duplex even at ionic strengths approaching the physiological conditions. With the aim to develop an optimized BioFET setup, the interaction kinetics between miR-155 and the chosen PNA was preliminarily investigated by using surface plasmon resonance (SPR). By exploiting both these results and our custom-made bioFET system, we were able to attain a low-cost, real-time, label-free and highly specific detection of miR-155 in the nano-molar range.
Insights
This study presents a novel method for detecting microRNAs (miRNAs) using peptide nucleic acid (PNA) probes and field-effect-transistor biosensors (bioFETs). This approach enables sensitive, real-time, and label-free detection of specific miRNAs like miR-155.
Area of Science:
- Biotechnology
- Molecular Biology
- Biosensor Technology
Background:
- MicroRNAs (miRNAs) are crucial gene regulators implicated in various diseases, including cancer.
- Altered miRNA expression necessitates sensitive clinical detection methods for biomarker applications.
- Field-effect-transistor-based biosensors (bioFETs) offer a promising platform for label-free miRNA quantification.
Purpose of the Study:
- To investigate the detection of human microRNA 155 (miR-155) using a novel peptide nucleic acid (PNA) probe.
- To optimize a bioFET setup for sensitive and specific miRNA detection.
- To evaluate the potential of PNA-bioFET systems for clinical applications.
Main Methods:
- Utilized a synthetic peptide nucleic acid (PNA) as a capturing probe for miR-155 detection.
- Employed Surface Plasmon Resonance (SPR) to study the interaction kinetics between miR-155 and the PNA probe.
- Developed and utilized a custom-made bioFET system for real-time, label-free miRNA detection.
Main Results:
- Demonstrated that PNA probes can form stable duplexes with target miRNAs under physiological conditions.
- Achieved sensitive detection of miR-155 in the nano-molar range using the PNA-bioFET system.
- Confirmed high specificity of the PNA-bioFET system for miR-155 detection.
Conclusions:
- The PNA-bioFET system provides a low-cost, real-time, and highly specific method for miR-155 detection.
- This technology holds significant potential for developing clinical diagnostic tools for miRNA-related diseases.
- Further optimization could lead to widespread adoption in clinical settings for miRNA biomarker analysis.
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