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Real-time in situ observation of P53-mediated cascade activation of apoptotic pathways with nucleic acid multicolor fluorescent probes based on symmetrical gold nanostars

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Abstract

T-2 toxin, one of the most dangerous natural pollutants, induces apoptosis through multiple pathways. Amongst, P53 mediated apoptosis pathway, an important collection of molecules, plays a key role in cell vital activity. Real-time monitoring of upstream and downstream activation relationships of P53 mRNA, Bax mRNA, and cytochrome c (Cyt c) in signaling pathways is of great significance for understanding the apoptotic machinery in human physiology. In this work, a novel nucleic acid multicolor fluorescent probe, based on silica-coated symmetric gold nanostars (S-AuNSs@SiO2), was developed for highly sensitive in situ real-time imaging of P53 mRNA, Bax mRNA, and Cyt c during T-2 toxin-induced apoptosis. The nucleic acid chains modified with carboxyl groups were modified on the surface of S-AuNSs@SiO2 by amide reaction. The complementary chains of targeted mRNA and the aptamer of targeted Cyt c were modified with different fluorophores, respectively, and successfully hybridized on S-AuNSs@SiO2 surface. When targets were present, the fluorescent chains bound to the targets and detached from the material, resulting in the quenched fluorescence being revived. The probes based on S-AuNSs showed excellent performance is partly ascribed to the presence of 20 symmetric “hot spots”. Notably, the amide-bonded probe exhibited excellent anti-interference capability against biological agents (nucleases and biothiols). During the real-time fluorescence imaging of T-2 toxin-induced apoptosis, the corresponding fluorescence signals of P53 mRNA, Bax mRNA, and Cyt c were observed sequentially. Therefore, S-AuNSs@SiO2 probe not only provides a novel tool for real-time monitoring of apoptosis pathways cascade but also has considerable potential in disease diagnosis and pharmaceutical medical.

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Acknowledgements

We are grateful for the financial support from the Jiangsu Agriculture Science and Technology Innovation Fund (No. CX (19)2005) and the Social Development Fund Project of Wuxi (No. N20201001).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Chenbiao Li, Peifang Chen, ** Wang

  2. School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Chenbiao Li, Peifang Chen, ** Wang

  3. International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, 214122, China

    Chenbiao Li, Peifang Chen, ** Wang

  4. Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi, 214122, China

    Chenbiao Li, Peifang Chen, ** Wang

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Real-time in situ observation of P53-mediated cascade activation of apoptotic pathways with nucleic acid multicolor fluorescent probes based on symmetrical gold nanostars

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Li, C., Chen, P., Ma, X. et al. Real-time in situ observation of P53-mediated cascade activation of apoptotic pathways with nucleic acid multicolor fluorescent probes based on symmetrical gold nanostars. Nano Res. 16, 5391–5400 (2023). https://doi.org/10.1007/s12274-022-5069-9

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