A Unique G-Quadruplex Aptamer: A Novel Approach for Cancer Cell Recognition, Cell Membrane Visualization, and RSV Infection Detection.
Chao-Da XiaoMing-Qing ZhongYue GaoZheng-Lin YangMeng-Hao JiaXiao-Hui HuYan XuXiang-Chun ShenPublished in: International journal of molecular sciences (2023)
Surface staining has emerged as a rapid technique for applying external stains to trace cellular identities in diverse populations. In this study, we developed a distinctive aptamer with selective binding to cell surface nucleolin (NCL), bypassing cytoplasmic internalization. Conjugation of the aptamer with a FAM group facilitated NCL visualization on live cell surfaces with laser confocal microscopy. To validate the aptamer-NCL interaction, we employed various methods, including the surface plasmon resonance, IHC-based flow cytometry, and electrophoretic mobility shift assay. The G-quadruplex formations created by aptamers were confirmed with a nuclear magnetic resonance and an electrophoretic mobility shift assay utilizing BG4, a G-quadruplex-specific antibody. Furthermore, the aptamer exhibited discriminatory potential in distinguishing between cancerous and normal cells using flow cytometry. Notably, it functioned as a dynamic probe, allowing real-time monitoring of heightened NCL expression triggered by a respiratory syncytial virus (RSV) on normal cell surfaces. This effect was subsequently counteracted with dsRNA transfection and suppressed the NCL expression; thus, emphasizing the dynamic attributes of the probe. These collective findings highlight the robust versatility of our aptamer as a powerful tool for imaging cell surfaces, holding promising implications for cancer cell identification and the detection of RSV infections.
Keyphrases
- flow cytometry
- respiratory syncytial virus
- label free
- gold nanoparticles
- sensitive detection
- loop mediated isothermal amplification
- magnetic nanoparticles
- magnetic resonance
- poor prognosis
- quantum dots
- single cell
- cell surface
- high throughput
- cell therapy
- high resolution
- biofilm formation
- induced apoptosis
- magnetic resonance imaging
- respiratory tract
- escherichia coli
- cell death
- climate change
- mesenchymal stem cells
- photodynamic therapy
- binding protein
- heavy metals