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Updated: Nov 17, 2025
Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay
Published on: June 9, 2017
Light up multiple protein dimers on cell surface based on proximity-induced fluorescence activation of DNA-templated
Lulu Xu1, Zheng Zhou2, Xiaolong Gou3
1Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, 400016, PR China; The Center for Clinical Molecular Medical Detection, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, PR China.
Abstract:
Efficient and multiple analysis of receptor protein dimers is highly necessary, due to their important role in the occurrence and development of cancer. Herein, we report a turn-on strategy to visualize human epidermal growth factor receptor (HER) dimers on cell surfaces. By taking advantages of specific aptamer recognition and proximity-induced fluorescence activation of DNA-templated sliver nanoclusters (DNA/AgNCs) by guanine (G)-rich sequence, we attached the two kind of DNA/AgNCs sequence with different fluorescence properties to the corresponding HER aptamer to form aptamer-functionalized AgNCs probes, and attached G-rich sequence to the corresponding HER aptamer as enhancer. In the presence of protein dimers, after aptamer specific recognition and binding, it will draw the dark AgNCs probes close to the G-rich probes and then excite corresponding fluorescence. As a result, this approach has successfully realized imaging of HER2:HER2 homodimer and HER2:HER3 heterodimer at the same time, which was provided a new idea for the simultaneous detection of multiple HER2 dimers in situ. This AgNCs-based light up strategy provides a potential tool for further investigation of protein dimerization on cell surface, which is more conducive to the mechanism research, accurate classification and treatment of cancer.
Insights
This study presents a new "turn-on" fluorescence method to visualize human epidermal growth factor receptor (HER) dimers on cancer cells. The technique enables simultaneous imaging of HER2 homodimers and HER2:HER3 heterodimers, aiding cancer research.
Area of Science:
- Biochemistry
- Molecular Biology
- Nanotechnology
Background:
- Receptor protein dimers play a crucial role in cancer development.
- Efficient analysis of these dimers is essential for understanding cancer.
- Current methods for dimer analysis may lack efficiency or multiplexing capabilities.
Purpose of the Study:
- To develop a novel
- turn-on
- fluorescence strategy for visualizing human epidermal growth factor receptor (HER) dimers on cell surfaces.
- To enable simultaneous detection and imaging of specific HER dimers, such as HER2:HER2 homodimers and HER2:HER3 heterodimers.
- To provide a new tool for cancer research, mechanism investigation, and classification.
Main Methods:
- Utilized aptamer recognition for specific binding to HER dimers.
- Employed DNA-templated silver nanoclusters (DNA/AgNCs) for fluorescence generation.
- Designed a proximity-induced fluorescence activation mechanism involving G-rich sequences.
- Developed aptamer-functionalized AgNCs probes and G-rich enhancer sequences.
Main Results:
- Successfully visualized HER2:HER2 homodimers on cell surfaces.
- Successfully visualized HER2:HER3 heterodimers on cell surfaces.
- Achieved simultaneous imaging of both HER2 homodimers and HER2:HER3 heterodimers in situ.
- Demonstrated a functional
- turn-on
- fluorescence system based on DNA/AgNCs proximity.
Conclusions:
- The developed AgNCs-based
- turn-on
- strategy offers a novel approach for visualizing and simultaneously detecting multiple HER dimers.
- This method provides a valuable tool for investigating protein dimerization on cell surfaces, crucial for cancer research.
- The strategy has potential applications in understanding cancer mechanisms, improving classification, and guiding treatment.
