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  1. Home
  3. Bioenergetic Shift And Proteomic Signature Induced By Lentiviral-transduction Of Gfp-based Biosensors.
  1. Home
  3. Bioenergetic Shift And Proteomic Signature Induced By Lentiviral-transduction Of Gfp-based Biosensors.

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Bioenergetic shift and proteomic signature induced by lentiviral-transduction of GFP-based biosensors.

Sarah Barakat1, Şeyma Çimen2, Seyed Mohammad Miri3

  • 1Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey.

Redox Biology
|November 7, 2024

View abstract on PubMed

Summary
This summary is machine-generated.

Green fluorescent protein (GFP) biosensors like HyPer7 can cause cellular stress and mitochondrial dysfunction due to hydrogen peroxide production. Researchers recommend caution and validation when using these fluorescent protein tools in biological studies.

Keywords:
Green fluorescent proteinsHyPer7 genetically encoded biosensorHydrogen peroxideOxidative stressProteomic signature

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Fluorescent proteins (FPs) are essential research tools but can cause cellular side effects.
  • Green fluorescent protein (GFP) maturation generates hydrogen peroxide (H₂O₂), potentially impairing cell function.
  • The impact of GFP-based biosensors on cellular homeostasis requires further investigation.

Purpose of the Study:

  • To evaluate the effects of the GFP-based HyPer7 biosensor on cellular homeostasis and proteome.
  • To identify potential risks associated with oxidative stress induced by HyPer7 expression.
  • To assess the reliability of FP-based tools in biological research.

Main Methods:

  • Utilized genome-integrated HyPer7 biosensor expression in cells.
  • Assessed mitochondrial membrane potential (MMP) and oxidative respiration.
  • Performed untargeted proteomics analysis to identify differentially expressed proteins.
  • Conducted functional annotation of identified proteins.

    • Main Results:

    • HyPer7 expression altered mitochondrial membrane potential (MMP), responsive to antioxidants and normoxia.
    • Significant impairment in mitochondrial oxidative respiration observed in HyPer7-expressing cells.
    • Proteomics identified 26 differentially expressed proteins linked to stress response and antioxidant mechanisms.

    Conclusions:

    • GFP-based biosensors, including HyPer7, can induce oxidative stress and mitochondrial dysfunction.
    • Cellular responses to FP expression warrant careful consideration and validation.
    • Ensuring comprehension of cellular responses enhances the reliability of FP-based research findings.