Department of Chemistry, University of Kurdistan, 66177-15175, Sanandaj, Iran.
Research Center for Nanotechnology, University of Kurdistan, 66177-15175, Sanandaj, Iran.
J Nanobiotechnology. 2022 Feb 8;20(1):73. doi: 10.1186/s12951-022-01274-3.
MicroRNAs have short sequences of 20 ~ 25-nucleotides which are similar among family members and play crucial regulatory roles in numerous biological processes, such as in cell development, metabolism, proliferation, differentiation, and apoptosis.
We reported a strategy for the construction of a dual-emission fluorescent sensor using carbon dots (CDs) and confirmed their applications for ratiometric microRNA-21 sensing and bioimaging of cancer cells in a microfluidic device. The composition of blue CDs (B-CDs) and yellow CDs (Y-CDs) depicts dual-emission behavior which is centered at 409 and 543 nm under an excitation wavelength of 360 nm. With increasing microRNA-21 concentration, the robust and specific binding of DNA probe functionalized B-CDs to complementary microRNA-21 target induced perturbations of probe structure and led to changing fluorescence intensity in both wavelengths. Consequently, the ratio of turn-on signal to turn-off signal is greatly altered. With monitoring of the inherent ratiometric fluorescence variation (ΔF/ΔF), as-prepared BY-CDs were established as an efficient platform for ratiometric fluorescent microRNA-21 sensing, with a wide linear range of 0.15 fM to 2.46 pM and a detection limit of 50 aM.
Furthermore, the proposed assay was applied for detecting microRNA-21 in dilute human serum samples with satisfactory recovery and also in MCF-7 cell lines in the range 3000 to 45,000 (cell mL) with a detection limit (3 cells in 10 μL), demonstrating the potential of the assay for clinic diagnosis of microRNA-associated disease. More importantly, the images revealed that MCF-7 cells well labeled with BY-CDs could exhibit the applicability of the proposed microfluidic system as an effective cell trapping device in bioimaging.
microRNAs 是一类具有 20-25 个核苷酸长度的短序列分子,在家族成员之间具有相似性,在许多生物过程中发挥着关键的调节作用,如细胞发育、代谢、增殖、分化和凋亡。
我们报道了一种使用碳点(CDs)构建双发射荧光传感器的策略,并在微流控装置中证实了其用于比率型 microRNA-21 传感和癌细胞的生物成像的应用。蓝色碳点(B-CDs)和黄色碳点(Y-CDs)的组成表现出双发射行为,在 360nm 激发波长下,中心发射波长分别为 409nm 和 543nm。随着 microRNA-21 浓度的增加,功能化 B-CDs 的 DNA 探针与互补 microRNA-21 靶标之间的强特异性结合导致探针结构的扰动,并导致两个波长的荧光强度发生变化。因此,开信号与关信号的比值发生了很大的变化。通过监测固有比率荧光变化(ΔF/ΔF),制备的 BY-CDs 被建立为一种有效的比率荧光 microRNA-21 传感平台,具有 0.15 fM 至 2.46 pM 的宽线性范围和 50 aM 的检测限。
此外,该测定法被用于检测稀释的人血清样品中的 microRNA-21,回收率令人满意,也被用于 MCF-7 细胞系(细胞 mL)的 3000 至 45000 范围内,检测限为 3 个细胞(10 μL),证明了该测定法在 microRNA 相关疾病临床诊断中的潜力。更重要的是,图像显示,用 BY-CDs 标记的 MCF-7 细胞能够表现出该微流控系统作为一种有效的细胞捕获装置在生物成像中的适用性。