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Updated: Oct 19, 2025
In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila
Published on: November 27, 2016
A switch-on molecular biosensor for detection of caspase-3 and imaging of apoptosis of cells
Rui Gong1,2, Dianbing Wang1, Ghulam Abbas1,2
1National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
Abstract:
Apoptosis is a form of programmed cell death that is essential for maintaining internal environmental stability. Disordered apoptosis can cause a variety of diseases; therefore, sensing apoptosis can provide help in study of mechanism of the relevant diseases and drug development. It is known that caspase-3 is a key enzyme involved in apoptosis and the expression of its activity is an indication of apoptosis. Here, we present a genetically encoded switch-on mNeonGreen2-based molecular biosensor. mNeonGreen2 is the brightest monomeric green fluorescent protein. The substrate of caspase-3, DEVD amino acid residues, is inserted in it, while cyclized by insertion of Nostoc punctiforme DnaE intein to abolish the fluorescence (inactive state). Caspase-3-catalyzed cleavage of DEVD linearizes mNeonGreen2 and rebuilds the natural barrel structure to restore the fluorescence (activated state). The characterization exhibited that the Caspase-3 biosensor has shortened response time, higher sensitivity, and prolonged functional shelf life in detection of caspase-3 amongst the existing counterparts. We also used the Caspase-3 biosensor to evaluate the effect of several drugs on the induction of apoptosis of HeLa and MCF-7 tumor cells and inhibition of Zika virus invasion.
Insights
Researchers developed a novel mNeonGreen2-based biosensor to detect caspase-3 activity, a key indicator of programmed cell death (apoptosis). This sensitive tool aids in disease research and drug development by enabling precise apoptosis monitoring.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Apoptosis, or programmed cell death, is crucial for health, and its dysregulation is linked to diseases.
- Caspase-3 activity is a key biomarker for apoptosis, making its detection vital for research and therapeutic development.
Purpose of the Study:
- To develop a genetically encoded, switch-on fluorescent biosensor for sensitive and real-time detection of caspase-3 activity.
- To characterize the biosensor's performance and demonstrate its utility in cellular models and drug screening.
Main Methods:
- Engineered a mNeonGreen2 fluorescent protein by inserting a caspase-3 substrate (DEVD) and an intein for fluorescence quenching.
- Designed the biosensor to regain fluorescence upon caspase-3-mediated cleavage, linearizing the protein.
- Validated the biosensor in HeLa and MCF-7 tumor cells to assess drug effects on apoptosis and Zika virus inhibition.
Main Results:
- The developed biosensor exhibits faster response times, enhanced sensitivity, and improved shelf-life compared to existing caspase-3 detection methods.
- Successfully utilized the biosensor to monitor drug-induced apoptosis in cancer cell lines.
- Demonstrated the biosensor's application in evaluating the inhibition of Zika virus invasion.
Conclusions:
- The novel mNeonGreen2-based biosensor provides a robust and efficient tool for monitoring apoptosis via caspase-3 activity.
- This biosensor has significant potential for advancing apoptosis research, disease mechanism studies, and drug discovery efforts.
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