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Glass Nanopipette-Based Plasmonic SERS Platform for Single-Cell MicroRNA-21 Sensing during Apoptosis
Yong Wang1,2, Dandan Wang3, Guohua Qi1
1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China.
Abstract:
As one of the most widely distributed microRNAs, microRNA-21 (miRNA-21) significantly regulates target genes' expression levels and participates in many cellular and intercellular activities, and its abnormal expression is always related to some diseases, especially cancer. Hence, detecting miRNA-21, as a biomarker, at the single-cell level helps us to reveal cell heterogeneity and expression level variation during the state change of cells. In this study, we constructed a gold nanoparticles nanomembrane (AuNPs-NM)-modified plasmonic glass nanopipette (P-nanopipette) surface-enhanced Raman scattering (SERS) sensing platform to sensitively detect content variation of the intracellular miRNA-21 during the electrostimulus (ES)-induced apoptosis process. The cytoplasm-located miRNA-21 was first extracted by using the extraction DNA (HP1)-modified P-nanopipette through a hybridization chain reaction (HCR). The nanopipette was then incubated with a labeling DNA (HP2) and reporter 4-MBA-modified Raman tag. The Raman signal (collected from the tip area near the orifice within 1 μm) showed a good response to the content variation of intracellular miRNA-21 under ES, and the proposed single-cell SERS detection platform provides a simple way to study intracellular substance change and evaluate cancer treatment outcomes.
Insights
This study introduces a novel nanopipette sensing platform for detecting microRNA-21 (miRNA-21) in single cells. The method sensitively tracks miRNA-21 changes during electrostimulus-induced apoptosis, aiding cancer research.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Nanotechnology
Background:
- MicroRNA-21 (miRNA-21) is a widely distributed microRNA involved in cellular processes and disease, particularly cancer.
- Abnormal miRNA-21 expression is linked to various diseases, making its detection crucial for understanding cell heterogeneity and state changes.
- Single-cell level detection of miRNA-21 is valuable for revealing variations during cellular events like apoptosis.
Purpose of the Study:
- To develop a sensitive platform for detecting intracellular miRNA-21 variations during electrostimulus-induced apoptosis at the single-cell level.
- To utilize a gold nanoparticles nanomembrane-modified plasmonic glass nanopipette with surface-enhanced Raman scattering (SERS) for this detection.
- To establish a method for studying intracellular substance changes and evaluating cancer treatment outcomes.
Main Methods:
- Construction of a gold nanoparticles nanomembrane (AuNPs-NM)-modified plasmonic glass nanopipette (P-nanopipette).
- Extraction of cytoplasm-located miRNA-21 using an extraction DNA (HP1)-modified P-nanopipette via a hybridization chain reaction (HCR).
- Incubation with a labeling DNA (HP2) and a 4-mercaptobenzoic acid (4-MBA)-modified Raman tag for SERS detection.
Main Results:
- The SERS sensing platform demonstrated sensitive detection of intracellular miRNA-21 content variation.
- A good response in Raman signal was observed correlating with miRNA-21 changes during electrostimulus (ES)-induced apoptosis.
- The method successfully monitored miRNA-21 dynamics at the single-cell level.
Conclusions:
- The developed P-nanopipette SERS platform offers a sensitive and simple approach for single-cell miRNA-21 detection.
- This platform can effectively study intracellular substance changes during cellular processes like apoptosis.
- The technology holds potential for evaluating cancer treatment outcomes by monitoring miRNA biomarkers.
