Quantification of MicroRNA in a Single Living Cell via Ionic Current Rectification-Based Nanopore for Triple Negative Breast Cancer Diagnosis,Analytical Chemistry

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Quantification of MicroRNA in a Single Living Cell via Ionic Current Rectification-Based Nanopore for Triple Negative Breast Cancer Diagnosis
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-04-23 , DOI: 10.1021/acs.analchem.3c05027
Shujie Zhang ₁ , Laibo Song ₁ , Ruina Zheng ₁ , Fang Zhang ₁ , Qimeng Wang ₁ , Xiaosui Mao ₁ , Jin-Xuan Fan ₁ , Bo Liu ₁ , Yuan-Di Zhao _{1,

2} , Wei Chen ₁

Affiliation

Accurate analysis of microRNAs (miRNAs) at the single-cell level is extremely important for deeply understanding their multiple and intricate biological functions. Despite some advancements in analyzing single-cell miRNAs, challenges such as intracellular interferences and insufficient detection limits still remain. In this work, an ultrasensitive nanopore sensor for quantitative single-cell miRNA-155 detection is constructed based on ionic current rectification (ICR) coupled with enzyme-free catalytic hairpin assembly (CHA). Benefiting from the enzyme-free CHA amplification strategy, the detection limit of the nanopore sensor for miRNA-155 reaches 10 fM and the nanopore sensor is more adaptable to complex intracellular environments. With the nanopore sensor, the concentration of miRNA-155 in living single cells is quantified to realize the early diagnosis of triple-negative breast cancer (TNBC). Furthermore, the nanopore sensor can be applied in screening anticancer drugs by tracking the expression level of miRNA-155. This work provides an adaptive and universal method for quantitatively analyzing intracellular miRNAs, which will greatly improve our understanding of cell heterogeneity and provide a more reliable scientific basis for exploring major diseases at the single-cell level.

中文翻译：

通过基于离子电流整流的纳米孔对单个活细胞中的 MicroRNA 进行定量，用于三阴性乳腺癌诊断

在单细胞水平上准确分析 microRNA (miRNA) 对于深入了解其多重且复杂的生物学功能极其重要。尽管在分析单细胞 miRNA 方面取得了一些进展，但细胞内干扰和检测限不足等挑战仍然存在。在这项工作中，基于离子电流整流（ICR）与无酶催化发夹组装（CHA）相结合，构建了一种用于定量单细胞miRNA-155检测的超灵敏纳米孔传感器。受益于无酶CHA扩增策略，纳米孔传感器对miRNA-155的检测限达到10 fM，纳米孔传感器对复杂的细胞内环境具有更强的适应性。利用纳米孔传感器，对活体单细胞中miRNA-155的浓度进行定量，实现三阴性乳腺癌（TNBC）的早期诊断。此外，纳米孔传感器还可以通过追踪miRNA-155的表达水平来筛选抗癌药物。这项工作为细胞内miRNA的定量分析提供了一种适应性强、通用的方法，将极大提高我们对细胞异质性的认识，为在单细胞水平上探索重大疾病提供更可靠的科学依据。

更新日期：2024-04-23

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