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首页> 外文期刊>Langmuir: The ACS Journal of Surfaces and Colloids >LRET-based biodetection of DNA release in live cells using surface-modified upconverting fluorescent nanoparticles
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LRET-based biodetection of DNA release in live cells using surface-modified upconverting fluorescent nanoparticles

机译：使用表面修饰的上转换荧光纳米粒子基于LRET的活细胞DNA释放生物检测

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摘要

In this work, we demonstrate near-infrared-to-visible upconverting fluorescent nanoparticles as a promising platform for lanthanide-based or luminescence resonance energy transfer (LRET)-based biodetection of DNA release in live cells. Highly monodispersed, stable aqueous suspension of nanoparticles, surface-functionalized with amino groups for binding to DNA, were prepared and characterized. These amino-functionalized nanoparticles were able to electrostatically bind to DNA and protect it from DNaseI degradation as shown by gel electrophoresis. Attachment of DNA to the nanoparticles was also confirmed by LRET, which was observed to occur between the donor nanoparticle and acceptor POPO-3 dye intercalating the DNA. The intrinsic fluorescence property of upconverting fluorescent nanoparticles makes them useful for tracking their cellular localization without the use of any additional fluorescent tag. We were able to track the movement of these DNA-loaded nanoparticles into the cell cytoplasm where they successfully released their genetic cargo. Successful transfection of the loaded DNA material in vitro and in vivo was confirmed by expression of its encoded green fluorescent protein (GFP) in Hela cells and induction of specific antibody in mice, respectively.
机译：在这项工作中，我们证明了近红外到可见光的上转换荧光纳米颗粒是基于镧系元素或基于发光共振能量转移（LRET）的活细胞DNA释放生物检测的有前途的平台。制备并表征了高度单分散，稳定的纳米颗粒水悬浮液，该表面经氨基表面官能化以与DNA结合。这些氨基官能化的纳米粒子能够与DNA静电结合，并保护其免受DNaseI降解（如凝胶电泳所示）。 LRET也证实了DNA与纳米颗粒的附着，观察到发生在供体纳米颗粒和嵌入DNA的受体POPO-3染料之间。上转换荧光纳米粒子的固有荧光特性使其可用于跟踪其细胞定位，而无需使用任何其他荧光标记。我们能够追踪到这些装载了DNA的纳米颗粒进入细胞质的过程，并在细胞质中成功释放了遗传物质。分别通过在Hela细胞中表达其编码的绿色荧光蛋白（GFP）和在小鼠中诱导特异性抗体，证实了体外和体内成功转染负载的DNA材料。

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