Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, PR China.
Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, PR China.
Anal Chim Acta. 2023 Apr 22;1251:341003. doi: 10.1016/j.aca.2023.341003. Epub 2023 Feb 22.
The development of DNA nanomachines provides a new strategy for the detection of tumor markers. In this work, an intelligent three-dimensional (3D) DNA walking machine with polynucleotide kinase (PNK) activator was designed, which was coupled with unique nanomachine formed by DNA nanowire cascade amplification reaction for versatile fluorescence detection of T4 PNK activity and messenger RNA (mRNA). When PNK exists, the free DNA walker was formed by hydrolysis cleavage of exonuclease, then the fluorophore-labeled report probe on the Au nanoparticles (NPs) was sheared during cycling cleavage reaction, thus the fluorescence signal was recovered for detection of PNK. Moreover, the DNA nanowires were produced by rolling ring amplification, then target mRNA sequentially initiated interval hybridization of hairpin probes through DNA nanowire, thus realizing DNA cascade reaction (DCR) with high "on" signal of DNA nanomachine for mRNA assay. This developed novel fluorescence nanomachine reported a new assay method with promising application for versatile targets and showed great potential for molecular-target therapies, and clinic diagnostics.
DNA 纳米机器的发展为肿瘤标志物的检测提供了新的策略。在这项工作中，设计了一种具有多核苷酸激酶 (PNK) 激活剂的智能三维 (3D) DNA 行走机，它与独特的纳米机器结合，通过 DNA 纳米线级联扩增反应进行 T4 PNK 活性和信使 RNA (mRNA) 的多功能荧光检测。当 PNK 存在时，通过核酸外切酶的水解裂解形成游离的 DNA 行走机，然后在循环裂解反应过程中剪切 Au 纳米粒子 (NPs) 上标记荧光探针的报告探针，从而恢复荧光信号以检测 PNK。此外，通过滚环扩增产生 DNA 纳米线，然后目标 mRNA 通过 DNA 纳米线依次引发发夹探针的间隔杂交，从而实现 DNA 级联反应 (DCR)，DNA 纳米机器的“开启”信号高，用于 mRNA 分析。该新型荧光纳米机器报道了一种具有广阔应用前景的多功能靶标新分析方法，为分子靶向治疗和临床诊断显示出巨大的潜力。
