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A Fluorescence-SERS Dual-Mode Nanoprobe for Imaging of HSP90 mRNA and Peroxynitrite in Living Cells
Yue He1, Ming-Jie Ye1, Cheng-Ye Xi1
1Key Laboratory for Advanced Materials, Shanghai Key Laboratory of Functional Materials Chemistry, Frontiers Science Center for Materiobiology & Dynamic Chemistry, School of Chemistry& Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China.
Abstract:
The dysregulation of heat shock protein 90 mRNA (HSP90 mRNA) and reactive oxygen species (ROS) is implicated in stress response and various diseases. Visualizing HSP90 mRNA and ROS dynamics is important to studying their interactions and related physiopathological mechanisms. However, effective methods for detecting both remain lacking. Herein, a covalent organic framework-based (COF-based) dual-mode nanoprobe is designed to monitor HSP90 mRNA and ONOO- (ROS model). The nanoprobe is prepared by in situ assembly of a COF shell as the aptamer carrier on the gold nanorods (AuNRs), followed by conjugation of the ONOO--responsive molecule, 4-mercaptophenylboronic acid (4-MPBA), to the AuNRs and modification of the HSP90 mRNA aptamer (HSP90MB) onto the COF shell. The prepared nanoprobe enables sensitive and selective fluorescence (FL) and surface-enhanced Raman spectroscopy (SERS) detection of HSP90 mRNA and ONOO-, respectively. The dual-channel detection highlights the advantages of facilitating spectral analysis and eliminating mutual interference. In addition, the proposed strategy visualizes a positive interaction between HSP90 mRNA and ONOO- in living cells, revealing their cellular response mechanism under stress conditions and related diseases.
Insights
A novel nanoprobe visualizes heat shock protein 90 mRNA (HSP90 mRNA) and reactive oxygen species (ROS) dynamics. This dual-mode detection advances understanding of cellular stress responses and related diseases.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Molecular Biology
Background:
- Heat shock protein 90 mRNA (HSP90 mRNA) and reactive oxygen species (ROS) dysregulation are linked to stress responses and diseases.
- Simultaneous visualization of HSP90 mRNA and ROS is crucial for understanding their interplay but current methods are limited.
Purpose of the Study:
- To develop a dual-mode nanoprobe for simultaneous detection of HSP90 mRNA and a reactive oxygen species model (peroxynitrite, ONOO⁻).
- To investigate the interaction between HSP90 mRNA and ONOO⁻ in living cells.
Main Methods:
- Fabrication of a covalent organic framework (COF)-based nanoprobe using gold nanorods (AuNRs) as a core.
- Functionalization of the nanoprobe with an ONOO⁻-responsive molecule (4-mercaptophenylboronic acid) and an HSP90 mRNA aptamer.
- Utilizing fluorescence (FL) and surface-enhanced Raman spectroscopy (SERS) for dual-channel detection.
Main Results:
- The COF-based nanoprobe achieved sensitive and selective detection of both HSP90 mRNA and ONOO⁻.
- Dual-channel detection minimized spectral interference and enhanced analysis.
- The study visualized a positive correlation between HSP90 mRNA and ONOO⁻ in living cells.
Conclusions:
- The developed dual-mode nanoprobe offers a powerful tool for studying molecular dynamics in cellular stress.
- This approach provides insights into the mechanisms underlying stress responses and related pathologies.
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