Dai Kaining, Zhao Jian, Li Lele, Fu Xiaojun
Sanbo Brain Hospital, Capital Medical University, Laboratory for Clinical Medicine, Capital Medical University, Beijing, 100070, China.
CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing, 100190, China.
Small Methods. 2025 Jun;9(6):e2401531. doi: 10.1002/smtd.202401531. Epub 2024 Nov 14.
Live-cell imaging of RNA in specific subcellular compartments is essential for elucidating the rich repertoire of cellular functions, but it has been limited by a lack of simple, precisely controlled methods. Here such an approach is presented via the combination of hybridization chain reaction and spatially restricted enzymatic activation with organelle-targeted delivery. The system can localize engineered DNA hairpins in the mitochondria, where target RNA-initiated chain reaction of hybridization events is selectively activated by a specific enzyme, enabling amplified RNA imaging with high precision. It is demonstrated that the approach is compatible with live cell visualization and enables the regulatable imaging of microRNA in mitochondria. Since in situ activation of the signal amplification with enzyme eliminates the need for genetically encoded protein overexpression, it is envisioned that this simple platform will be broadly applicable for precise RNA imaging with subcellular resolution in a variety of biological processes.
对特定亚细胞区室中的RNA进行活细胞成像对于阐明丰富的细胞功能至关重要，但一直受到缺乏简单、精确可控方法的限制。在此，通过杂交链式反应与细胞器靶向递送的空间受限酶促激活相结合，提出了一种这样的方法。该系统可以将工程化的DNA发夹定位在线粒体中，在那里，目标RNA引发的杂交事件链式反应被一种特定的酶选择性激活，从而实现高精度的RNA扩增成像。结果表明，该方法与活细胞可视化兼容，并能够对线粒体中的微小RNA进行可调节成像。由于用酶原位激活信号放大无需基因编码蛋白的过表达，因此可以设想，这个简单的平台将广泛适用于在各种生物学过程中以亚细胞分辨率进行精确的RNA成像。
