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Ligand-Induced Self-Complementing Tag (LiSC-Tag) as an Epitope Tag for Live-Cell Super-Resolution Imaging and
Liuxin Tu1, Hong Qin1, Siyuan Luo1
1School of Pharmaceutical Sciences, Tsinghua University, Beijing 100084, China.
Journal of the American Chemical Society
|September 18, 2025
Summary
Researchers developed a new Ligand-induced Self-complementing Tag (LiSC-tag) for advanced live-cell super-resolution microscopy. This novel epitope tag improves protein labeling specificity and efficiency for clearer intracellular protein imaging and manipulation.
Area of Science:
- Biophysics
- Molecular Biology
- Cell Biology
Background:
- Epitope tags are valuable for live-cell super-resolution imaging due to their small size.
- Current epitope tags often suffer from insufficient labeling specificity and efficiency for intracellular proteins.
- Existing methods struggle to achieve high-resolution imaging of endogenous proteins in live cells.
Purpose of the Study:
- To develop a novel epitope tag for enhanced live-cell super-resolution imaging.
- To improve labeling specificity and efficiency for intracellular protein visualization.
- To enable functional manipulation of cellular proteins using advanced imaging techniques.
Main Methods:
- Development of the Ligand-induced Self-complementing Tag (LiSC-tag) based on a split FKBP mutant.
- Utilizing small-molecule fluorescent probes for ligand-induced self-complementation.
- Application of LiSC-tag in multicolor nanoscopy techniques including STED, STORM, and SIM.
- Engineering tandem arrays of LiSC-tags to amplify fluorescence signals.
Main Results:
- The LiSC-tag achieved high labeling specificity and efficiency in live-cell super-resolution imaging.
- Demonstrated multicolor nanoscopy imaging capabilities using STED, STORM, and SIM.
- Observed superior brightness and photostability compared to split fluorescent proteins.
- Successfully imaged endogenous proteins and manipulated protein localization and degradation.
Conclusions:
- The LiSC-tag represents a significant advancement for live-cell super-resolution microscopy.
- It offers a versatile tool for both high-resolution imaging and functional studies of cellular proteins.
- LiSC-tag technology facilitates multicolor imaging and protein manipulation, overcoming previous limitations.
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