State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China.
J Am Chem Soc. 2021 Sep 8;143(35):14394-14401. doi: 10.1021/jacs.1c07719. Epub 2021 Aug 25.
Genetically encoded molecular tools are crucial for live cell RNA imaging, and few are available for endogenous RNA imaging. We develop a new genetically encoded sensor using conformation switching RNA induced fluorogenic proteins that enable multicolor and signal-amplified imaging of endogenous RNAs. The sensor system is designed with an RNA sensing module and a degron-fused fluorescent protein reporter. Target RNA induces conformation switching of the RNA sensing module to form RNA aptamers that stabilize the degron-fused protein for fluorogenic imaging. This sensor is demonstrated for high-contrast imaging of mRNA abundance and dynamics in live cells. Moreover, the sensor system is extended to a multicolor palette by screening fluorogenic proteins of distinct colors, and engineered into a signal amplifier using the split fluorescent protein design. The sensor is further exploited for imaging lncRNA MALAT-1 and its translocation dynamics during mitosis. Our sensor system can afford a valuable platform for RNA imaging in biomedical research and clinical theranostics.
基因编码的分子工具对于活细胞 RNA 成像至关重要，但可用于内源性 RNA 成像的工具却很少。我们开发了一种新的基因编码传感器，该传感器使用构象转换 RNA 诱导的荧光蛋白，能够对内源性 RNA 进行多色和信号放大成像。该传感器系统设计有一个 RNA 感应模块和一个融合降解元件的荧光蛋白报告子。靶 RNA 诱导 RNA 感应模块的构象转换，形成 RNA 适体，稳定融合降解元件的蛋白，用于荧光成像。该传感器可用于高对比度地成像活细胞中 mRNA 的丰度和动力学。此外，该传感器系统通过筛选不同颜色的荧光蛋白进行了多色扩展，并使用分裂荧光蛋白设计将其工程化为信号放大器。该传感器还用于成像 lncRNA MALAT-1 及其在有丝分裂期间的转位动力学。我们的传感器系统可为生物医学研究和临床治疗中的 RNA 成像提供有价值的平台。
