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Chemoselective Modification of Viral Surfaces via Bioorthogonal Click Chemistry
Published on: August 19, 2012
inCu-click: Enabling copper-catalyzed click chemistry inside living cells
Keqing Nian1, Hillary Ho2, Arnon Chuenrudeemol3
1Department of Bioengineering, Northeastern University, Boston, MA, United States.
Methods in Enzymology
|June 25, 2026
Summary
A new inCu-click platform enables efficient intracellular bioconjugation using copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) in live cells. This method minimizes cell toxicity by localizing copper ions, allowing robust labeling of biomolecules with high cell viability.
Area of Science:
- Bioconjugation Chemistry
- Cellular Imaging
- Molecular Biology
Background:
- Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is a key bioconjugation technique.
- Intracellular CuAAC labeling in live cells is challenging due to copper toxicity.
- Existing methods often require high copper concentrations, leading to cell death.
Purpose of the Study:
- To develop a novel platform for efficient intracellular CuAAC labeling in live cells.
- To overcome the limitations of high copper concentrations and cell toxicity.
- To enable real-time visualization of intracellular biomolecular dynamics.
Main Methods:
- Development of the inCu-click platform using a DNA-conjugated ligand to localize copper ions.
- Liposomal delivery of the inCu-click ligand into cells.
- Application of the platform for fluorescent labeling of phospholipids and alkyne drugs in live cells.
Main Results:
- The inCu-click system facilitates efficient CuAAC under reduced intracellular copper levels.
- Robust fluorescent labeling of nascent phospholipids and alkyne drugs was achieved.
- Minimal impact on cell viability was observed during intracellular labeling.
Conclusions:
- The inCu-click platform offers a practical solution for intracellular click chemistry in live cells.
- This technology enables reliable visualization of biomolecular dynamics in complex cellular environments.
- The platform demonstrates broad applicability for various intracellular labeling applications.
