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Related Experiment Video

Updated: Sep 13, 2025

Monitoring of Ubiquitin-proteasome Activity in Living Cells Using a Degron dgn-destabilized Green Fluorescent Protein GFP-based Reporter Protein

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Published on: November 10, 2012

Tracking Intracellular Labile Iron with a Genetically Encoded Fluorescent Reporter System Based on Protein Stability.

Ali Akyol¹, Şeyma Çimen², Benjamin Gottschalk¹

¹Gottfried Schatz Research Center, Molecular Biology and Biochemistry, Medical University of Graz, Neue Stiftingtalstraße 6, 8010 Graz, Austria.

|August 1, 2025

Summary

Researchers developed IronFist, a new tool for tracking iron (Fe2+) in live cells. This reporter reveals how cells handle iron, showing significant differences in response to iron supplements and highlighting cell-to-cell variations in iron levels.

Keywords:

Hr domain fluorescence microscopy iron carbohydrate nanoparticles iron reporter iron-dependent degron labile iron pool

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Published on: June 18, 2020

Area of Science:

Cell Biology
Biochemistry
Molecular Biology

Background:

Cellular iron homeostasis is crucial for numerous biological processes.
Dysregulation of iron levels is linked to various diseases, including neurodegenerative disorders and cancer.
Accurate methods for dynamic tracking of intracellular labile iron are needed.

Purpose of the Study:

To develop and validate a novel genetically encoded fluorescent reporter, IronFist, for real-time monitoring of labile ferrous ions (Fe2+) in live cells.
To investigate cellular responses to different iron sources using the IronFist reporter.
To assess cell-to-cell variability in iron handling.

Main Methods:

Development of a bicistronic fluorescent reporter system (IronFist) utilizing an iron-sensitive hemerythrin-like (Hr) domain fused to mNeonGreen and a reference fluorescent protein (mCherry).
Utilizing IronFist for endpoint measurements and time-lapse imaging to track labile iron dynamics.
Treating cells with iron(II) sulfate and iron carbohydrate nanoparticles to observe cellular iron pool fluctuations.

Main Results:

IronFist successfully enables dynamic tracking of labile ferrous ions (Fe2+) in live cells.
Most cells exhibit low basal labile iron levels, but significant increases are observed upon iron supplementation.
Differential cellular responses to iron(II) sulfate (faster, stronger) versus iron carbohydrate nanoparticles (slower, weaker) were detected, alongside substantial cell-to-cell heterogeneity.

Conclusions:

IronFist provides a valuable tool for assessing cellular iron homeostasis and understanding iron-related pathologies.
The reporter enables high-content, single-cell level tracking of iron dynamics, filling a critical methodological gap.
Findings highlight the complex and heterogeneous nature of cellular iron handling.