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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Related Experiment Video

Updated: Jan 3, 2026

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
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Engineering Photoactivatability in Genetically Encoded Voltage and pH Indicators.

Sungmoo Lee1, Yoon-Kyu Song2,3, Bradley J Baker1,4

  • 1Center for Functional Connectomics, Brain Science Institute, Korea Institute of Science and Technology, Seoul, South Korea.

Frontiers in Cellular Neuroscience
|November 19, 2019
PubMed
Summary

Researchers developed photoactivatable genetically encoded voltage (GEVI) and pH (GEPI) indicators. This innovation overcomes background fluorescence, enabling precise optical monitoring of neuronal activity and intracellular pH changes.

Keywords:
GEVIPA-Bongwoori-R3PA-GFPPA-ecliptic pHluorinecliptic pHluorinpH sensorphotoactivatablevoltage indicator

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Biophysics

Background:

  • Genetically encoded indicators allow optical monitoring of neuronal activity in specific cell populations.
  • High background fluorescence often hinders signal detection in optical monitoring.
  • Photoactivatable indicators, like sPA-GCaMP6f, limit fluorescence to defined regions of interest, improving signal-to-noise ratio.

Purpose of the Study:

  • To develop photoactivatable genetically encoded voltage indicators (GEVIs) and pH indicators (GEPIs).
  • To adapt the photoactivation strategy, previously used for calcium indicators, to voltage and pH sensing.
  • To overcome limitations of background fluorescence in optical neuroimaging.

Main Methods:

  • Investigated three photoactivatable GEVI candidates: one using a circularly-permuted fluorescent protein, another employing Förster resonance energy transfer (FRET), and a third based on ecliptic pHluorin.
  • Engineered a pH-sensitive photoactivatable GFP variant.
  • Evaluated photoactivation properties and voltage-dependent fluorescence changes of the developed indicators.

Main Results:

  • Identified a GEVI candidate utilizing ecliptic pHluorin that demonstrated successful photoactivation.
  • Observed voltage-dependent fluorescence changes in the ecliptic pHluorin-based GEVI.
  • Developed a novel pH-sensitive photoactivatable GFP that responds to intracellular pH fluctuations.

Conclusions:

  • Photoactivation strategies can be successfully applied to genetically encoded voltage and pH indicators.
  • The ecliptic pHluorin-based GEVI offers a promising tool for targeted optical monitoring of neuronal voltage.
  • A new photoactivatable pH indicator provides a means for optical measurement of intracellular pH dynamics.