Detection of extracellular RNAs in cancer and viral infection via tethered cationic lipoplex nanoparticles containing molecular beacons.
Detection of extracellular RNAs in cancer and viral infection via tethered cationic lipoplex nanoparticles containing molecular beacons.
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DOI：
10.1021/ac401983w
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发表时间：
2013-12-03
影响因子：
7.4
通讯作者：
Lee, L. James
中科院分区：
文献类型：
--
作者：
Wu, Yun;Kwak, Kwang Joo;Agarwal, Kitty;Marras, Alexander;Wang, Chao;Mao, Yicheng;Huang, Xiaomeng;Ma, Junyu;Yu, Bo;Lee, Robert;Vachani, Anil;Marcucci, Guido;Byrd, John C.;Muthusamy, Natarajan;Otterson, Gregory;Huang, Kun;Castro, Carlos E.;Paulaitis, Michael;Nana-Sinkam, Serge P.;Lee, L. James
非侵入性早期检测方法有可能降低癌症和传染病的死亡率。在这里，我们提出了一种新的检测方法，通过该方法，含有分子信标（MB）的拴系阳离子lipoplex纳米颗粒可以捕获癌细胞来源的外泌体或病毒，并在一个步骤中识别封装的RNA。首先使用一系列超离心和Exoquick™分离试剂盒分别从细胞培养基和人血清中分离外来体。连接到薄玻璃板表面上的阳离子脂质复合物纳米颗粒通过静电相互作用捕获带负电荷的病毒或细胞分泌的外泌体，以形成更大的纳米级复合物。脂质复合物/病毒或脂质复合物/外来体融合导致病毒/外来体RNA和MB在脂质复合物内混合。在靶RNA特异性结合MB后，通过MB的荧光信号容易地鉴定富含靶RNA的外泌体。在不稀释样品的情况下原位检测靶细胞外RNA导致现有方法（例如qRT-PCR）无法实现的高检测灵敏度。在这里，我们使用慢病毒和肺癌患者的血清来证明这一概念。
Non-invasive early detection methods have the potential to reduce mortality rates of both cancer and infectious diseases. Here, we present a novel assay by which tethered cationic lipoplex nanoparticles containing molecular beacons (MBs) can capture cancer cell-derived exosomes or viruses, and identify encapsulated RNAs in a single step. A series of ultracentrifugation and Exoquick™ isolation kit were first used to isolate exosomes from the cell culture medium and human serum respectively. Cationic lipoplex nanoparticles linked onto the surface of a thin glass plate capture negatively charged viruses or cell-secreted exosomes by electrostatic interactions to form larger nanoscale complexes. Lipoplex/virus or lipoplex/exosome fusion leads to the mixing of viral/exosomal RNAs and MBs within the lipoplexes. After the target RNAs specially bind to the MBs, exosomes enriched in target RNAs are readily identified by the fluorescence signals of MBs. The in situ detection of target extracellular RNAs without diluting the samples leads to high detection sensitivity not achievable by existing methods, e.g. qRT-PCR. Here we demonstrate this concept using lentivirus and serum from lung cancer patients.
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DOI：
10.1038/nrc2961
发表时间：
2010-12
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1073/pnas.0804549105
发表时间：
2008-07-29
影响因子：
11.1
作者：
Mitchell, Patrick S.;Parkin, Rachael K.;Tewari, Muneesh
通讯作者：
Tewari, Muneesh
血清中 microRNA 的表征：一类用于诊断癌症和其他疾病的新型生物标志物
影响因子：
44.1
作者：
Chen, Xi;Ba, Yi;Zhang, Chen-Yu
通讯作者：
Zhang, Chen-Yu
影响因子：
2.1
作者：
McGillivray, Duncan J.;Valincius, Gintaras;Losche, Mathias
通讯作者：
Losche, Mathias
影响因子：
20.3
作者：
Eisfeld, Ann-Kathrin;Marcucci, Guido;Bloomfield, Clara D.
通讯作者：
Bloomfield, Clara D.