Application of the FRET method for monitoring the dynamics of caspase-3 activation during apoptosis in living cells

Tongsheng Chen; Da Xing

doi:10.1117/12.572753

18 January 2005 Application of the FRET method for monitoring the dynamics of caspase-3 activation during apoptosis in living cells

Tongsheng Chen, Da Xing

Author Affiliations +

Proceedings Volume 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II; (2005) https://doi.org/10.1117/12.572753
Event: Photonics Asia, 2004, Beijing, China

Abstract

Activation of caspase-3 is a central event in apoptosis. A fluorescence techniques, fluorescence resonance energy transfer (FRET), was used to study the dynamic of caspase-3 activation during apoptosis induced by tumor necrosis factor TNF-α in living cells. The FRET probe consists a CFP (cyan fluorescent protein) and a Venus (YFP mutant, yellow fluorescent protein) with a specialized linker containing the caspase-3 cleavage sequence: DEVD (Luo et al., 2001). Human lung adenocarcinoma cell line (ASTC-a-1) were stably expressed with the FRET probe and then were treated by TNF-α, respectively. Experimental results showed that FRET could monitor more insensitively the dynamic of caspase-3 activation in real-time in vivo, and this technique will be highly useful for correlating the caspase-3 activation with other apoptotic events and for rapid-screening of potential drugs that may target the apoptotic process.

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Tongsheng Chen and Da Xing "Application of the FRET method for monitoring the dynamics of caspase-3 activation during apoptosis in living cells", Proc. SPIE 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II, (18 January 2005); https://doi.org/10.1117/12.572753

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KEYWORDS

Fluorescence resonance energy transfer

Cell death

Luminescence

Venus

Fluorescence correlation spectroscopy

Fluorescent proteins

Proteins

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