Confined Target-Triggered Hot Spots for In Situ SERS Analysis of Intranuclear Genotoxic Markers

化学 遗传毒性 原位 拉曼光谱 纳米技术 DNA 生物物理学 生物化学 光学 材料科学 毒性 物理 有机化学 生物
作者
Lingfei Han,Yingjiu Zhou,Zheng Tan,Hongtan Zhu,Yexin Hu,Xiao Ma,Feng Zheng,Feng Feng,Chen Wang,Wenyuan Liu
出处
期刊:Analytical Chemistry [American Chemical Society]
卷期号:95 (15): 6312-6322 被引量:3
标识
DOI:10.1021/acs.analchem.2c05147
摘要

The γH2AX is a type of confined target in nuclei which is highly expressed around the damaged DNA during genotoxicity and has therefore been identified as a marker of genotoxicity. Convenient and intuitive in situ real-time detection of γH2AX is crucial for an accurate assessment of genotoxicity. Selective and nondestructive surface-enhanced Raman spectroscopy (SERS) is suitable to achieve this goal. However, the detection of substances in the nucleus by SERS is still limited due to the contradiction of probes between the nuclei entry efficiency and signal enhancement. This study utilized the characteristics of γH2AX as a confined target and constructed a γH2AX immunosensor based on gold nanoprobes with a small size (15 nm), which was modified with the TAT nuclear targeting peptide to ensure high nuclei entry efficiency. Once DNA damage was induced, the local overexpression of γH2AX further recruited the probe through immune recognition, so that hot spots could be assembled in situ to generate strong Raman signals, which were applied to evaluate the genotoxicity of drug impurities. This study proposed a novel SERS detection strategy, characterized by confined target-induced size conversion and hot spot formation, for in situ real-time analysis of intranuclear targets at the single-living-cell level, which intelligently simplified the structure of SERS probes and the operation process.
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