Fluorescence Resonance Energy Transfer-Based DNA Tetrahedron Nanotweezer for Highly Reliable Detection of Tumor-Related mRNA in Living Cells.
Fluorescence Resonance Energy Transfer-Based DNA Tetrahedron Nanotweezer for Highly Reliable Detection of Tumor-Related mRNA in Living Cells.
复制标题
DOI：
10.1021/acsnano.7b00725
复制
发表时间：
2017-04-25
期刊：
影响因子：
17.1
通讯作者：
Tan W
中科院分区：
文献类型：
--
作者：
He L;Lu DQ;Liang H;Xie S;Luo C;Hu M;Xu L;Zhang X;Tan W
活细胞中肿瘤相关mRNA的准确检测和成像对于早期癌症检测具有很大的希望。然而，目前，大多数设计用于成像细胞内mRNA的探针面临复杂生物基质产生的内在干扰，并导致不可避免的假阳性信号。为了解决这个问题，开发了一种称为DNA四面体纳米镊子（DTNT）的细胞内DNA纳米探针，以基于FRET（荧光共振能量转移）“关”到“开”信号读出模式来可靠地对活细胞中的肿瘤相关mRNA进行成像。DTNT由四个单链DNA自组装而成。在不存在靶mRNA的情况下，分别标记的供体和受体荧光团被分离，从而诱导低FRET效率。然而，在靶mRNA的存在下，DTNT将其结构从开放状态改变为闭合状态，从而使双荧光团紧密接近以获得高FRET效率。DTNT具有良好的细胞渗透性、快速响应性和良好的生物相容性。此外，细胞内成像实验表明，DTNT可以有效地区分癌细胞和正常细胞，此外，区分活细胞中mRNA表达水平的变化。DTNT纳米探针也表现出最小的影响，探针浓度，分布和激光功率为其他比率探针。更重要的是，由于FRET“关”到“开”信号读出模式，DTNT纳米探针几乎完全避免了由于内在干扰（例如核酸酶消化、蛋白质结合和复杂生物基质中的热力学波动）而导致的假阳性信号。这一设计蓝图可应用于开发功能强大的DNA纳米机器，用于生物医学研究和临床早期诊断。
Accurate detection and imaging of tumor-related mRNA in living cells hold great promise for early cancer detection. However, currently, most probes designed to image intracellular mRNA confront intrinsic interferences arising from complex biological matrices and resulting in inevitable false-positive signals. To circumvent this problem, an intracellular DNA nanoprobe, termed DNA tetrahedron nanotweezer (DTNT), was developed to reliably image tumor-related mRNA in living cells based on the FRET (fluorescence resonance energy transfer) “off” to “on” signal readout mode. DTNT was self-assembled from four single-stranded DNAs. In the absence of target mRNA, the respectively labeled donor and acceptor fluorophores are separated, thus inducing low FRET efficiency. However, in the presence of target mRNA, DTNT alters its structure from the open to closed state, thus bringing the dual fluorophores into close proximity for high FRET efficiency. The DTNT exhibited high cellular permeability, fast response and excellent biocompatibility. Moreover, intracellular imaging experiments showed that DTNT could effectively distinguish cancer cells from normal cells and, moreover, distinguish among changes of mRNA expression levels in living cells. The DTNT nanoprobe also exhibits minimal effect of probe concentration, distribution and laser power as other ratiometric probe. More importantly, as a result of the FRET “off” to “on” signal readout mode, the DTNT nanoprobe almost entirely avoids false-positive signals due to intrinsic interferences, such as nuclease digestion, protein binding and thermodynamic fluctuations in complex biological matrices. This design blueprint can be applied to the development of powerful DNA nanomachines for biomedical research and clinical early diagnosis.
登录
查看更多内容
影响因子：
14.9
作者：
Chen AK;Behlke MA;Tsourkas A
通讯作者：
Tsourkas A
影响因子：
15
作者：
Jayagopal, Ashwath;Halfpenny, Kristin C.;Perez, Jonas W.;Wright, David W.
通讯作者：
Wright, David W.
DOI：
10.1073/pnas.2233244100
发表时间：
2003-11-11
影响因子：
11.1
作者：
Bratu, DP;Cha, BJ;Tyagi, S
通讯作者：
Tyagi, S
影响因子：
7.4
作者：
Medley, CD;Drake, TJ;Tan, WH
通讯作者：
Tan, WH
影响因子：
15
作者：
Li, Juan;Zheng, Cheng;Cansiz, Sena;Wu, Cuichen;Xu, Jiehua;Cui, Cheng;Liu, Yuan;Hou, Weijia;Wang, Yanyue;Zhang, Liqin;Teng, I-ting;Yang, Huang-Hao;Tan, Weihong
通讯作者：
Tan, Weihong