An intramolecular catalytic hairpin assembly on a DNA tetrahedron for mRNA imaging in living cells: improving reaction kinetics and signal stability.
An intramolecular catalytic hairpin assembly on a DNA tetrahedron for mRNA imaging in living cells: improving reaction kinetics and signal stability.
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DNA四面体上的分子内催化发夹组装用于活细胞中的mRNA成像：改善反应动力学和信号稳定性
DOI：
10.1039/c9sc04916a
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发表时间：
2019-12-18
期刊：
影响因子：
8.4
通讯作者：
Yang R
中科院分区：
文献类型：
--
作者：
Qing Z;Hu J;Xu J;Zou Z;Lei Y;Qing T;Yang R
基于动态DNA自组装（DDSA）的无酶扩增技术最近被开发用于活细胞中mRNA的原位检测。然而，传统DDSA放大器的信号产生主要依赖于解离探针在体溶液中的随机扩散，通常伴随着动力学差和复杂生物系统的干扰。在这项工作中，提出了一种基于分子内催化发夹组件（intra-CHA）设计的新型放大器，用于活细胞中mRNA的FRET成像。与自由催化发夹组装（free- cha）相比，cha内探针H1和H2同时固定在DNA四面体上。两者之间的距离更近，理论上cha内H1和H2的局部浓度比自由cha高约808倍，初始反应速率提高了15.6倍。由于空间约束效应，靶催化信号生成的反应动力学得到了显著改善。由于cha内放大器的三维纳米结构，H1和H2无需任何转染或纳米载体进入细胞，探针及其产物不受生物干扰，为活细胞中mRNA的可靠成像提供了更高的信号稳定性。分子内催化发夹组件在DNA四面体上实现mRNA在活细胞中的成像。空间约束效应可以加速靶触发信号的产生，具有优异的细胞通透性和FRET信号稳定性。
Enzyme-free amplification techniques based on dynamic DNA self-assembly (DDSA) have recently been developed for the in situ detection of mRNA in living cells. However, signal generation in traditional DDSA amplifiers is mainly dependent on the random diffusion of dissociative probes in a bulk solution, which is generally accompanied by poor kinetics and interference from complex biological systems. In this work, a new amplifier based on the design of an intramolecular catalytic hairpin assembly (intra-CHA) is proposed for the FRET imaging of mRNA in living cells. Compared with that in the free catalytic hairpin assembly (free-CHA), probes H1 and H2 in intra-CHA were simultaneously fixed on a DNA tetrahedron. The distance between them was closer, the local concentration of H1 and H2 in intra-CHA was theoretically approximately 808-times higher than that in free-CHA, and the initial reaction rate was enhanced 15.6 fold. Due to the spatial confinement effect, the reaction kinetics for target-catalyzed signal generation were significantly improved. By virtue of the three-dimensional nanostructure, H1 and H2 in the intra-CHA amplifier entered cells without any transfection or nanocarrier, and the probes and their products were free from biological interference, providing much higher signal stability for the reliable imaging of mRNA in living cells. An intramolecular catalytic hairpin assembly is implemented on a DNA tetrahedron for mRNA imaging in living cells. The spatial confinement effect enables the acceleration of target-triggered signal generation, with excellent cell permeability and FRET signal stability.
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影响因子：
15
作者：
He L;Lu D;Liang H;Xie S;Zhang X;Liu Q;Yuan Q;Tan W
通讯作者：
Tan W
影响因子：
15
作者：
Karunanayake Mudiyanselage APKK;Yu Q;Leon-Duque MA;Zhao B;Wu R;You M
通讯作者：
You M
DOI：
10.1073/pnas.0407024101
发表时间：
2004-10-26
影响因子：
11.1
作者：
Dirks, RM;Pierce, NA
通讯作者：
Pierce, NA
影响因子：
15
作者：
Cheglakov, Zoya;Cronin, Timothy M.;Weizmann, Yossi
通讯作者：
Weizmann, Yossi
影响因子：
7.4
作者：
Qing, Zhihe;He, Xiaoxiao;He, Dinggeng
通讯作者：
He, Dinggeng