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Peptide-Fluorophores/AuNP conjugate-based Two-Photon Excited Fluorescent nanosensor for Caspase-3 Activity Imaging Assay in Living Cells and Tissue

Abstract

Caspase-3-mediated cell apoptosis is one of the major reasons for hepatic ischemia reperfusion injury (HIRI) in liver surgery. Thus, the measurement of caspase-3 activity in liver cells or tissue might be used as a window of opportunity to explore the mechanism of HIRI and screen new protective drugs. Hence in the present study, by taking advantage of the excellent optical properties of the two-photon fluorescent dye (TPdye) BMVC (Ethyl-4-[3, 6-Bis (1-methyl-4-vinylpyridium iodine)-9H-carbazol-9-yl)]) such as the relative weak fluorescence emission at its free molecular format and great enhanced fluorescence emission when it is inserted into double-stranded DNA (dsDNA), we designed a novel fluorescence resonance energy transfer (FRET) strategy-based two-photon absorption (TPA) nanosensor via the assembly of the TPdye-labeled peptide (CANLNDEVDK-BMVC) onto AuNP's surface for measurement of caspase-3 activity in live cells and IR-treated Rat liver tissues. The AuNP/peptide-TPdyes nanosensor was initially in "off" state due to the efficiently fluorescence emission quenching of AuNPs to the adjacent TPdyes BMVC, while the fluorescence emission will be recovered when the BMVC molecule was released from the AuNP's surface and further great enhanced when it was inserted into dsDNA. Successful caspase-3 activities assay in apoptotic cells and Rat liver tissue slices reveal that this AuNP/peptide-TPdyes nanoconjugate not only is a robust, sensitive, and selective sensor for in vitro caspase-3 assay but also can be efficiently delivered into live cells as well as tissues and act as a “signal-on” fluorescent biosensor for specific, high-contrast imaging assay of enzymatic activities. Moreover, our design also provides a methodology model scheme for development of future AuNP/TPdye-based two-photon fluorescent probes for in vitro or in vivo determination of biological or biologically relevant species.

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Publication details

The article was received on 11 Apr 2017, accepted on 11 May 2017 and first published on 12 May 2017


Article type: Research Article
DOI: 10.1039/C7MD00177K
Citation: Med. Chem. Commun., 2017, Accepted Manuscript
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    Peptide-Fluorophores/AuNP conjugate-based Two-Photon Excited Fluorescent nanosensor for Caspase-3 Activity Imaging Assay in Living Cells and Tissue

    Q. Ge, N. Wang, J. Li and R. Yang, Med. Chem. Commun., 2017, Accepted Manuscript , DOI: 10.1039/C7MD00177K

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