Shanghai Key Lab of Chemical Assessment and Sustainability, School of Chemical Science and Engineering, College of Environmental Science and Technology, Tongji University, Shanghai, 200092, China; Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA; Department of Chemistry, Tsinghua University, Beijing, 100084, PR China.
College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou, 450001, PR China; Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
Anal Chim Acta. 2017 Aug 29;983:1-8. doi: 10.1016/j.aca.2017.07.017. Epub 2017 Jul 14.
Simultaneous detection and imaging of multiple intracellular messenger RNA (mRNAs) hold great significant for early cancer diagnostics and preventive medicine development. Herein, we propose a multiple-targeted graphene oxide (GO) nanocarrier that can simultaneously detect and image different type mRNAs in living cells. First of all, in vitro detection of multiple targets have been realized successfully based on the multiple-targeted GO nanocarrier with linear relationship ranging from 3 nM to 200 nM, as well as sensitive detection limit of 1.84 nM for manganese superoxide dismutase (Mn-SOD) mRNA and 2.45 nM for β-actin mRNA. Additionally, this nanosensing platform composed of fluorescent labelled single strand DNA probes and GO nanocarrier can identify Mn-SOD mRNA and endogenous mRNA of β-actin in living cancer cells, showing rapid response, high specificity, nuclease stability, and good biocompatibility during the cell imaging. Thirdly, changes of the expression levels of mRNA in living cells before or after the drug treatment can be monitored successfully. By using multiple ssDNA as probes and GO nanocarrier as the cellular delivery cargo, the proposed simultaneous multiple-targeted sensing platform will be of great potential as a powerful tool for intracellular trafficking process from basic research to clinical diagnosis.
同时检测和成像多个细胞内信使 RNA(mRNA)对于癌症早期诊断和预防医学的发展具有重要意义。在此,我们提出了一种多靶向氧化石墨烯(GO)纳米载体,可用于在活细胞中同时检测和成像不同类型的 mRNA。首先,基于多靶向 GO 纳米载体成功实现了对多个靶标的体外检测,线性范围为 3 nM 至 200 nM,以及对锰超氧化物歧化酶(Mn-SOD)mRNA 的检测限为 1.84 nM,β-肌动蛋白 mRNA 的检测限为 2.45 nM。此外,由荧光标记的单链 DNA 探针和 GO 纳米载体组成的纳米传感平台可在活癌细胞中识别 Mn-SOD mRNA 和内源性 β-肌动蛋白 mRNA,在细胞成像过程中表现出快速响应、高特异性、核酸酶稳定性和良好的生物相容性。第三,成功监测了药物处理前后活细胞中 mRNA 表达水平的变化。通过使用多个 ssDNA 作为探针和 GO 纳米载体作为细胞递药载体,所提出的同时多靶向传感平台将成为从基础研究到临床诊断的细胞内运输过程的有力工具。