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. 2021 Sep 9;65(3):540–549. doi: 10.1007/s11427-021-1986-7

A switch-on molecular biosensor for detection of caspase-3 and imaging of apoptosis of cells

Rui Gong 1,2, Dianbing Wang 1, Ghulam Abbas 1,2, Shimin Li 1,2, Qian Liu 1, Mengmeng Cui 1, Xian-En Zhang 1,2,3,✉
PMCID: PMC8449214  PMID: 34536207

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.

Supporting Information

The supporting information is available online at 10.1007/s11427-021-1986-7. The supporting materials are published as submitted, without typesetting or editing. The responsibility for scientific accuracy and content remains entirely with the authors.

Keywords: apoptosis, caspase-3, molecular biosensor, molecular imaging, Zika virus

Electronic supplementary material

11427_2021_1986_MOESM1_ESM.docx (1.2MB, docx)

Supplementary material, approximately 1.17 MB.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (21890743), National Key Research and Development Program of China (2017YFA0205500, 2018YFA0902702), the Strategic Priority Research Program of the Chinese Academy of Sciences, China (XDB29050100). We thank Yu Hou and Liu Song for their helpful discussions.

Compliance and ethics The author(s) declare that they have no conflict of interest.

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Supplementary Materials

11427_2021_1986_MOESM1_ESM.docx (1.2MB, docx)

Supplementary material, approximately 1.17 MB.

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