State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China.
Angew Chem Int Ed Engl. 2021 Jun 7;60(24):13244-13250. doi: 10.1002/anie.202014798. Epub 2021 Feb 2.
New tools for single-cell interrogation enable deeper understanding of cellular heterogeneity and associated cellular behaviors and functions. Information of RNA expression in single cell could contribute to our knowledge of the genetic regulatory circuits and molecular mechanism of disease development. Although significant progresses have been made for intracellular RNA analysis, existing methods have a trade-off between operational complexity and practical feasibility. We address this challenge by combining the ionic current rectification property of nanopipette reactor with duplex-specific nuclease-assisted hybridization chain reaction for signal amplification to realize a simple and practical intracellular nanosensor with minimal invasiveness, which enables single-cell collection and electrochemical detection of intracellular RNA with cell-context preservation. Systematic studies on differentiation of oncogenic miR-10b expression levels in non-malignant breast cells, metastatic breast cancer cells as well as non-metastatic breast cancer cells were then realized by this nanotool accompanied by assessment of different drugs effects. This work has unveiled the ability of electrochemistry to probe intracellular RNA and opened new opportunities to study the gene expression and heterogeneous complexity under physiological conditions down to single-cell level.
单细胞检测新技术使人们能够更深入地了解细胞异质性及其相关的细胞行为和功能。单细胞内的 RNA 表达信息有助于我们了解遗传调控回路和疾病发展的分子机制。尽管在细胞内 RNA 分析方面已经取得了重大进展，但现有方法在操作复杂性和实际可行性之间存在权衡。我们通过将纳米管反应器的离子电流整流特性与双链特异性核酸酶辅助杂交链式反应相结合，实现了信号放大，从而实现了一种简单实用的细胞内纳米传感器，具有最小的侵入性，能够在保留细胞背景的情况下进行单细胞内 RNA 的收集和电化学检测。通过该纳米工具，我们系统地研究了致癌 miR-10b 表达水平在非恶性乳腺细胞、转移性乳腺癌细胞和非转移性乳腺癌细胞中的分化情况，并评估了不同药物的作用。这项工作揭示了电化学探测细胞内 RNA 的能力，并为在生理条件下研究基因表达和单细胞水平的异质性复杂性开辟了新的机会。
