<br>由内源性 5'-三磷酸腺苷驱动和加速的荧光发光 3D DNA Walker，用于活细胞中灵敏、快速的无标记 MicroRNA 检测和成像,Analytical Chemistry

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由内源性 5'-三磷酸腺苷驱动和加速的荧光发光 3D DNA Walker，用于活细胞中灵敏、快速的无标记 MicroRNA 检测和成像
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-05-20 , DOI: 10.1021/acs.analchem.4c00664
Hongmin Lei ₁ , Jie Zhou ₁ , Fang Liu ₁ , Yichen Han ₁ , Yaqin Chai ₁ , Ruo Yuan ₁

Affiliation

在此，荧光发光 3D DNA 步行器 (FLDW) 由内源性 5'-三磷酸 (ATP) 分子提供动力和加速，构建了一种生物传感器，用于灵敏、快速、无标记地检测和成像 microRNA-221 (miRNA- 221）在恶性肿瘤细胞中。令人印象深刻的是，ATP作为FLDW的驱动力和加速器可以显着加快FLDW的运行速度，减少信号错误的可能性，并提高检测和成像的灵敏度。当由目标 miRNA-221 转化的输出 DNA H1-op 启动 FLDW 时，锚定到 AuNP 的 S 链中富含 G 的序列暴露形成 G-四链体 (G4s)，硫黄素 T (ThT) 嵌入G4s发出强烈的荧光，实现对目标miRNA-221的灵敏、快速检测。同时，利用 ThT 与 G4 的特异性结合和弱背景荧光响应，可以直接且经济有效地提高无标记测定的信噪比。所提出的FLDW系统通过采用催化发夹组装（CHA）来提高靶标的转化，可以实现对1 pM至10 nM范围内的靶标miRNA-221的灵敏检测，检测限为0.19 pM。此外，通过利用肿瘤微环境中存在的丰富 ATP，FLDW 实现了癌细胞中 miRNA-221 的快速准确成像。该策略为癌细胞中生物标志物的检测和成像提供了一种创新且高速的无标记方法，有望成为人类疾病生物分析、诊断和预后的强大工具。

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A Fluorescence Light-Up 3D DNA Walker Driven and Accelerated by Endogenous Adenosine-5′-triphosphate for Sensitive and Rapid Label-Free MicroRNA Detection and Imaging in Living Cells

Herein, a fluorescence light-up 3D DNA walker (FLDW) was powered and accelerated by endogenous adenosine-5′-triphosphate (ATP) molecules to construct a biosensor for sensitive and rapid label-free detection and imaging of microRNA-221 (miRNA-221) in malignant tumor cells. Impressively, ATP as the driving force and accelerator for FLDW could significantly accelerate the operation rate of FLDW, reduce the likelihood of errors in signaling, and improve the sensitivity of detection and imaging. When FLDW was initiated by output DNA H1-op transformed by target miRNA-221, G-rich sequences in the S strand, anchored to AuNP, were exposed to form G-quadruplexes (G4s), and thioflavin T (ThT) embedded in the G4s emitted intense fluorescence to realize sensitive and rapid detection of target miRNA-221. Meanwhile, the specific binding of ThT to G4 with a weak background fluorescence response was utilized to enhance the signal-to-noise ratio of the label-free assay straightforwardly and cost-effectively. The proposed FLDW system could realize sensitive detection of the target miRNA-221 in the range of 1 pM to 10 nM with a detection limit of 0.19 pM by employing catalytic hairpin assembly (CHA) to improve the conversion of the target. Furthermore, by harnessing the abundant ATP present in the tumor microenvironment, FLDW achieved rapid and accurate imaging of miRNA-221 in cancer cells. This strategy provides an innovative and high-speed label-free approach for the detection and imaging of biomarkers in cancer cells and is expected to be a powerful tool for bioanalysis, diagnosis, and prognosis of human diseases.

更新日期：2024-05-20

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