Determination of microRNA-451a (miRNA-451a) by a Reduced Graphene Oxide (rGO)-Based Fluorescence Assay with Duplex-Specific Nuclease (DSN)
Determination of microRNA-451a (miRNA-451a) by a Reduced Graphene Oxide (rGO)-Based Fluorescence Assay with Duplex-Specific Nuclease (DSN)
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DOI：
10.1080/00032719.2023.2232485
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发表时间：
2023-07-01
影响因子：
2
通讯作者：
Liu，Bin
中科院分区：
文献类型：
--
作者：
Tao，Xueqing;Qin，Yan;Liu，Bin
MIR-451a是一种与乳腺癌选择性雌激素受体调节剂治疗过程中耐药发展密切相关的生物分子。准确、灵敏地监测其在细胞内的水平，对肿瘤的药物选择和评价具有重要意义。然而，通常可以获得的单链DNA探针不能有效地进入细胞，通过避免核酸酶消化而灵敏地发出可靠的信号。在此，报道了一种检测活细胞中miR-451a的传感系统，该系统具有高特异性和灵敏度。该体系由还原的氧化石墨烯(RGO)、双链特异性核酸酶(DSN)和荧光标记的单链DNA纳米探针组成。RGO表面的荧光探针提供了超低的背景和对内切酶消化的抵抗力。双链特异性核酸酶依次触发了单链DNA纳米探针的杂交循环和裂解。在最佳条件下，信号放大提供了1 FM miR-451a的检测下限，线性范围从1 FM到100 nM。最后，将所建立的方法用于乳腺癌组织中miR-451a的检测。这一策略为miR-451a相关肿瘤的临床诊断和评估提供了一种有前景的工具。
MiR-451a is a biomolecule tightly related to resistance development during treatment with selective estrogen-receptor modulators for breast cancer. Accurately and sensitively monitoring its level in cells is of great significance for the drug selection and the evaluation of this cancer. However, the commonly obtainable ssDNA probe cannot efficiently enter the cell to sensitively emit reliable signals by avoiding nuclease digestion. Herein, a sensing system is reported to monitor miR-451a in living cells with high specificity and sensitivity. The system consisted of reduced graphene oxide (rGO), duplex-specific nuclease (DSN), and a fluorescence-labeled ssDNA nanoprobe. The fluorescence probe on the rGO surface provided an ultra-low background and resistance to endonuclease digestion. Duplex-specific nuclease sequentially triggered the hybridization cycle and the cleavage of ssDNA nanoprobe. The signal amplification provided a detection limit of 1 fM miR-451a using the optimal conditions with a linear range from 1 fM to 100 nM. Finally, the developed method was used to determine miR-451a in breast cancer tissue. This strategy provides a promising tool for clinical diagnosis and evaluation of miR-451a related cancer.