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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Updated: Jan 18, 2026

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

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Visualizing NEK9 in action: aptamer-based fluorescent probes for real-time live-cell imaging.

Liyu Zhang¹, Ying Yang¹, Lidangzhi Mo¹

¹Shaanxi Institute for Pediatric Diseases, Xi'an Children's Hospital, Xi'an, China.

Nucleosides, Nucleotides & Nucleic Acids

|September 10, 2025

Summary

Researchers developed a novel aptamer-based fluorescent probe for live-cell imaging of NEK9 kinase. This tool overcomes limitations of traditional methods, enabling clear visualization of NEK9 dynamics with minimal cell toxicity.

Keywords:

Aptamer NEK9 capture-SELEX fluorescent probe platform live-cell imaging

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Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

Published on: May 17, 2010

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Last Updated: Jan 18, 2026

Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

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Published on: March 16, 2017

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

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Published on: August 6, 2014

Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging

Published on: May 17, 2010

Area of Science:

Molecular Biology
Biochemistry
Cell Biology

Background:

Live-cell imaging is crucial for understanding protein dynamics in biological research and medicine.
Conventional labeling techniques face challenges like poor probe permeability and cytotoxicity, hindering effective live-cell imaging.

Purpose of the Study:

To develop and validate a novel aptamer-based fluorescent probe for real-time imaging of NEK9 kinase in living cells.
To overcome the limitations of existing live-cell imaging methods.

Main Methods:

Identified a DNA aptamer (Apt-011) that selectively binds NEK9 using in vitro capture-SELEX.
Engineered a 'signal-on' aptamer-based fluorescent probe utilizing aptamer-induced conformational changes to separate fluorophore-quencher pairs.
Validated the probe's performance in live-cell imaging, assessing NEK9 visualization and cytotoxicity.

Main Results:

Successfully identified Apt-011, a DNA aptamer with high specificity for NEK9.
Developed a novel 'signal-on' aptamer-based fluorescent probe platform for NEK9 imaging.
Demonstrated successful visualization of intracellular NEK9 in live cells with minimal cytotoxicity (cell viability > 95%).

Conclusions:

The developed aptamer-based fluorescent probe provides a robust tool for studying NEK9 kinase and its associated signaling pathways.
This work establishes a generalizable strategy for overcoming key bottlenecks in live-cell imaging through rational aptamer engineering.