Imaging of caspase-3 activation by a novel FRET probe composed of CFP and DsRed

Juquiang Lin; Zhihong Zhang; Bifeng Liu; Qingming Luo

doi:10.1117/12.667181

2 February 2006 Imaging of caspase-3 activation by a novel FRET probe composed of CFP and DsRed

Juquiang Lin, Zhihong Zhang, Bifeng Liu, Qingming Luo

Proceedings Volume 6026, ICO20: Biomedical Optics; 60260V (2006) https://doi.org/10.1117/12.667181
Event: ICO20:Optical Devices and Instruments, 2005, Changchun, China

Abstract

Caspases-3 is a kind of cysteine proteases and plays an important role in cell apoptosis. It has been reported that caspase-3 activation can be real-time detected in living cells by fluorescence resonance energy transfer (FRET) between an enhanced cyan fluorescent protein and enhanced yellow fluorescent protein. However, the large spectral overlap between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) emission and the highly sensitivity to pH of YFP restricted their detecting sensitivity and reliability. CFP and red fluorescent protein (DsRed) possess superb wavelength separation of donor and acceptor emission spectra and DsRed was insensitive to pH, so the FRET probe composed of CFP and DsRed would be more suitable for imaging caspase-3 activation than the FRET probe composed of CFP and YFP. We constructed a vector that encoded CRS (caspase-3 recognition site) fused with CFP and DsRed (CFP-CRS-DsRed). In CFP-CRS-DsRed expressing tumor cells, FRET from CFP to DsRed could be detected. In the Clinical applications of cancer chemotherapy, cisplatin is one of the most broadly used drugs. It was already confirmed that caspase-3 was activated in HeLa cell treated by cisplatin. When the cells were stimulated with cisplatin, we found that the FRET efficient was remarkably decreased and then disappeared. It indicated that actived caspase-3 cleaved the CFP-CRS-DsRed fusion protein at CRS site. Thus, the FRET probe of CFP-CRS-DsRed could sensitively and reliably monitor caspase-3 activation in living cell. This probe will be highly useful for rapid-screening potential drugs that may target the apoptotic process and for imaging tumors in vivo.

Citation Download Citation

Juquiang Lin, Zhihong Zhang, Bifeng Liu, and Qingming Luo "Imaging of caspase-3 activation by a novel FRET probe composed of CFP and DsRed", Proc. SPIE 6026, ICO20: Biomedical Optics, 60260V (2 February 2006); https://doi.org/10.1117/12.667181

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KEYWORDS

Fluorescence resonance energy transfer

Cell death

Fluorescent proteins

Proteins

Luminescence

In vivo imaging

Chromium

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