Department of Chemistry, Shanghai Stomatological Hospital, State Key Lab of Molecular Engineering of Polymers, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Institutes of Biomedical Sciences Fudan University, Shanghai 200433, China.
Anal Chem. 2022 Mar 8;94(9):4030-4038. doi: 10.1021/acs.analchem.1c05223. Epub 2022 Feb 25.
Spatial imaging of RNAs in single cells is extremely charming for deciphering of regulatory mechanisms in multiple migration modes during tumor metastasis. Herein, enzyme-free-mediated cascade amplified nanoprobes were designed for in situ single-molecule imaging of dual-microRNAs (miRNAs) in switchable migrating cells. Differential expression and localization of dual-miRNAs were clearly exhibited in multiple cell lines attributed to enhanced sensitivity via the cascade signal amplification strategy. Significantly, in situ three-dimensional (3D) imaging of dual-miRNAs in transition of cell migration phenotypes was successfully reconstructed in both non-confined and confined microenvironments in vitro, of which differential spatial distribution was observed in a single cell. This is very promising for exploring key roles of spatial RNA distribution in migrating cells at the single-molecule level, which will advance revealing the molecular mechanism and physical principle in 3D cell migration in vivo.
在单细胞中对 RNA 进行空间成像对于破译肿瘤转移过程中多种迁移模式下的调控机制极具吸引力。在此，设计了无酶介导的级联放大纳米探针，用于可切换迁移细胞中单分子水平的双 microRNAs（miRNAs）的原位成像。通过级联信号放大策略，增强了灵敏度，从而在多个细胞系中清晰地显示了双 miRNAs 的差异表达和定位。值得注意的是，在体外非约束和约束微环境中成功地重建了细胞迁移表型转变过程中双 miRNAs 的原位三维（3D）成像，在单个细胞中观察到了差异的空间分布。这对于探索单细胞中迁移细胞中空间 RNA 分布在关键作用非常有前景，这将推进对体内 3D 细胞迁移的分子机制和物理原理的揭示。
