Optical Control of Metal Ion Probes in Cells and Zebrafish Using Highly Selective DNAzymes Conjugated to Upconversion Nanoparticles.
Optical Control of Metal Ion Probes in Cells and Zebrafish Using Highly Selective DNAzymes Conjugated to Upconversion Nanoparticles.
复制标题
DOI：
10.1021/jacs.8b09867
复制
发表时间：
2018-11
影响因子：
15
通讯作者：
Zhenglin Yang;K. Y. Loh;Yu Chu;Ruopei Feng;Nitya Sai Reddy Satyavolu;Mengyi Xiong;Stephanie M Nakamata Huynh-Stephan
中科院分区：
文献类型：
--
作者：
Zhenglin Yang;K. Y. Loh;Yu Chu;Ruopei Feng;Nitya Sai Reddy Satyavolu;Mengyi Xiong;Stephanie M Nakamata Huynh-Stephan
金属离子在体外和体内的时空分布对于我们理解金属离子在生物系统中的作用至关重要，然而具有高空间和时间分辨率的探测金属离子的方法数量非常有限，特别是在体内。为了克服这一局限性，我们报告了一种Zn 2+特异性近红外（NIR）DNAzyme纳米探针，用于实时金属离子跟踪与时空控制在早期胚胎和斑马鱼幼虫。通过将光笼化的DNA酶缀合到镧系元素掺杂的上转换纳米颗粒（UCNPs）上，我们已经实现了深层组织穿透近红外980 nm光到365 nm发射的上转换。然后，紫外光光子有效地光解腺苷核糖核苷酸2 '-OH上含有硝基苄基的底物链，从而允许含有Zn 2+选择性DNA酶的互补DNA链进行酶促切割。含有最初被BHQ 1和Dabcyl猝灭剂猝灭的可见FAM荧光团的产物在切割后释放，导致更高的荧光信号。DNAzyme-UCNP探针通过在活细胞和斑马鱼胚胎中的NIR生物成像窗口中激发并在可见光区检测来实现Zn 2+传感。在这项研究中，我们介绍了一个平台，可用于了解时空控制的Zn 2+分布，从而深入了解细胞内和体内模型中的动态Zn 2+离子分布。
Spatial and temporal distributions of metal ions in vitro and in vivo are crucial in our understanding of the roles of metal ions in biological systems, and yet there is a very limited number of methods to probe metal ions with high space and time resolution, especially in vivo. To overcome this limitation, we report a Zn2+-specific near-infrared (NIR) DNAzyme nanoprobe for real-time metal ion tracking with spatiotemporal control in early embryos and larvae of zebrafish. By conjugating photocaged DNAzymes onto lanthanide-doped upconversion nanoparticles (UCNPs), we have achieved upconversion of a deep tissue penetrating NIR 980 nm light into 365 nm emission. The UV photon then efficiently photodecages a substrate strand containing a nitrobenzyl group at the 2'-OH of adenosine ribonucleotide, allowing enzymatic cleavage by a complementary DNA strand containing a Zn2+-selective DNAzyme. The product containing a visible FAM fluorophore that is initially quenched by BHQ1 and Dabcyl quenchers is released after cleavage, resulting in higher fluorescent signals. The DNAzyme-UCNP probe enables Zn2+ sensing by exciting in the NIR biological imaging window in both living cells and zebrafish embryos and detecting in the visible region. In this study, we introduce a platform that can be used to understand the Zn2+ distribution with spatiotemporal control, thereby giving insights into the dynamical Zn2+ ion distribution in intracellular and in vivo models.