Xu Tianqi, Li Fan, Jin Yichao, Jia Feng, Wang Yu, Lv Tao, Zhang Yueyue, Li Min, Fan Chunhai, Li Mingqiang, Zhang Xiaohua, Wang Shaopeng, Zuo Xiaolei
Institute of Molecular Medicine, Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, Department of Neurosurgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200127, China.
State Key Laboratory of Synergistic Chem-Bio Synthesis, School of Chemistry and Chemical Engineering, New Cornerstone Science Laboratory, Frontiers Science Center for Transformative Molecules, Zhangjiang Institute for Advanced Study and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai, 200240, China.
Angew Chem Int Ed Engl. 2025 Aug 25;64(35):e202506640. doi: 10.1002/anie.202506640. Epub 2025 Jul 9.
The transient release of metal ions at neuronal cell membranes holds significant implications in neurophysiology and pathology. Consequently, there is a pressing need for sensitive methodologies capable of quantitatively mapping local metal ion concentrations in live cells, aiming to unravel potential mechanisms underlying neurological disorders. DNAzymes have emerged as versatile tools for metal ion detection in live cells, owing to their specificity and other inherent advantages. Efforts to broaden their utility have focused on enhancing the stability and sensitivity of DNAzymes. Inspired by the beneficial effects of molecular chaperones on the stability and functionality of proteins, we present in this study the development of tetrahedral DNA frameworks (TDF) as chaperones for DNAzymes, aiming at enhancing the stability and activity of DNAzymes. Integration of TDF with DNAzymes is shown to significantly enhance metal ion detection performance, resulting in elevated stability and a two-fold increase in sensitivity, attributed to alterations in the local net charge induced by TDF. In vitro investigations demonstrate that the nanodevice developed here faithfully maps Cu concentrations within a range of 0-10 µM on the membrane of neuronal cells, meeting the requirements for Cu sensing under both physiological and pathological conditions. This work presents an easily adaptable approach to enhance the performance of DNAzymes and lays the foundations for the development of other DNAzyme-based sensitive detection nanodevices.
金属离子在神经元细胞膜上的瞬时释放对神经生理学和病理学具有重要意义。因此，迫切需要能够定量绘制活细胞中局部金属离子浓度的灵敏方法，以揭示神经系统疾病潜在的机制。由于其特异性和其他固有优势，脱氧核酶已成为活细胞中金属离子检测的通用工具。拓展其应用的努力主要集中在提高脱氧核酶的稳定性和灵敏度上。受分子伴侣对蛋白质稳定性和功能有益影响的启发，我们在本研究中提出开发四面体DNA框架（TDF）作为脱氧核酶的伴侣，旨在提高脱氧核酶的稳定性和活性。TDF与脱氧核酶的整合显示出显著提高金属离子检测性能，稳定性提高，灵敏度提高两倍，这归因于TDF诱导的局部净电荷变化。体外研究表明，这里开发的纳米装置能够准确绘制神经元细胞膜上0-10µM范围内的铜浓度，满足生理和病理条件下铜传感的要求。这项工作提出了一种易于应用的方法来提高脱氧核酶的性能，并为开发其他基于脱氧核酶的灵敏检测纳米装置奠定了基础。
