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Optokinetically Encoded Nanoprobe-Based Multiplexing Strategy for MicroRNA Profiling

机译:基于光动力学编码的基于纳米探针的MicroRNA分析多路复用策略

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摘要

Multiplexed real-time analysis on multiple interacting molecules and particles is needed to obtain information on binding patterns between multiple ligands and receptors, specificity of bond formations, and interacting pairs in a complex medium, often found in chemical and biological systems, and difference in binding affinity and kinetics for different binding pairs in one solution. In particular, multiplexed profiling of microRNA (miRNA) in a reliable, quantitative manner is of great demand for the use of miRNA in cell biology, biosensing, and clinical diagnostic applications, and accurate diagnosis of cancers with miRNA is not possible without detecting multiple miRNA sequences in a highly specific manner. Here, we report a multiplexed molecular detection strategy with optokinetically (OK) coded nanoprobes (NPs) that show high photostability, distinct optical signals, and dynamic behaviors on a supported lipid bilayer (SLB) (OK-NLB assay). Metal NPs with three distinct dark-field light scattering signals [red (R), green (G), and blue (B)] and three different target miRNA half-complements were tethered to a two dimensionally fluidic SLB with mobile (M) or immobile (I) state. In situ single-particle monitoring and normalized RGB analysis of the optokinetically combinatorial assemblies among three M-NPs and three I-NPs with dark-field microscopy (DFM) allow for differentiating and quantifying 9 different miRNA targets in one sample. The OK-NP-based assay enables simultaneous detection of multiple miRNA targets in a highly quantitative, specific manner within 1 h and can be potentially used for diagnosis of different cancer types. We validated the OK-NLB assay with single-base mismatched experiments and HeLa cell-extracted total RNA samples by comparing the assay results to the quantitative reverse transcription polymerase chain reaction (qRT-PCR) results.
机译:需要对多个相互作用的分子和颗粒进行多路实时分析,以获得有关多个配体和受体之间的结合模式,键形成的特异性以及复杂介质中相互作用对的信息(通常在化学和生物系统中发现)以及结合差异一种溶液中不同结合对的亲和力和动力学。特别是,以可靠,定量的方式对microRNA(miRNA)进行多重分析对在细胞生物学,生物传感和临床诊断应用中使用miRNA具有很高的需求,并且如果不检测多个miRNA,就不可能用miRNA准确诊断癌症。以高度特定的方式进行排序。在这里,我们报告了一种具有光动力学(OK)编码的纳米探针(NP)的多重分子检测策略,该策略在支持的脂质双层(SLB)上显示出高的光稳定性,独特的光信号和动态行为(OK-NLB分析)。金属NP具有三个不同的暗场光散射信号[红色(R),绿色(G)和蓝色(B)]和三个不同的目标miRNA半补体被束缚到带有可移动(M)或可移动的二维流体SLB静止(I)状态。利用暗场显微镜(DFM)对三个M-NP和三个I-NP之间的光动力学组合进行原位单粒子监测和标准化RGB分析,可区分和定量一个样品中的9种不同miRNA靶标。基于OK-NP的测定法能够在1小时内以高度定量,特异的方式同时检测多个miRNA靶标,可潜在地用于诊断不同类型的癌症。通过将测定结果与定量逆转录聚合酶链反应(qRT-PCR)结果进行比较,我们通过单碱基错配实验和HeLa细胞提取的总RNA样品验证了OK-NLB测定。

著录项

  • 来源
    《Journal of the American Chemical Society》 |2017年第9期|3558-3566|共9页
  • 作者单位

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

    Department of Chemistry, Seoul National University, Seoul 08826, South Korea;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
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  • 入库时间 2022-08-18 03:07:56

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