Chen Zhenyin, Chen Wei, Xu Cun, Song Haozhi, Ji Xin, Jiang Haodong, Duan Hongtao, Li Zehao, Gao Wankai, Yao Tuoxin, Zhang Zhongxuan, He Liuqin, Yin Yulong, Yang Nanyang, Tian Wenjing, Wu Jiahui, Li Xing
Interdisciplinary Science Center, State Key Laboratory of Integrated Management of Pest Insects and Rodents,Institute of Zoology, Chinese Academy of Sciences, 100101, Beijing, China.
Department of Pulmonary and Critical Care Medicine, the Affiliated Hospital of Southwest Medical University, 646000, Luzhou, Sichuan, China.
Nat Commun. 2025 Jan 9;16(1):518. doi: 10.1038/s41467-024-55093-1.
Fluorogenic RNA aptamers have various applications, including use as fluorescent tags for imaging RNA trafficking and as indicators of RNA-based sensors that exhibit fluorescence upon binding small-molecule fluorophores in living cells. Current fluorogenic RNA:fluorophore complexes typically emit visible fluorescence. However, it is challenging to develop fluorogenic RNA with near-infrared (NIR) fluorescence for in vivo imaging and sensing studies. To address this issue, we identify and modulate red fluorescent protein-like fluorophores to bind Squash, a highly folded fluorogenic RNA. One of these fluorophores, DFQL-1T, exhibits photostable NIR fluorescence when bound to Squash, enabling RNA visualization in living mammalian cells and mice. With Squash:DFQL-1T complexes, we generate RNA-based sensors for detecting non-coding RNAs and small molecule targets in living mammalian cells and in mice. These studies reveal a fluorogenic RNA:fluorophore complex that can be readily developed into NIR fluorescent RNA tags for in vivo imaging and sensing.
荧光RNA适配体有多种应用，包括用作成像RNA转运的荧光标签以及作为基于RNA的传感器的指示剂，这些传感器在活细胞中与小分子荧光团结合时会发出荧光。当前的荧光RNA：荧光团复合物通常发出可见荧光。然而，开发具有近红外（NIR）荧光的荧光RNA用于体内成像和传感研究具有挑战性。为了解决这个问题，我们鉴定并调节红色荧光蛋白样荧光团以结合南瓜素（Squash），一种高度折叠的荧光RNA。这些荧光团之一，DFQL-1T，与南瓜素结合时表现出光稳定的近红外荧光，能够在活的哺乳动物细胞和小鼠中实现RNA可视化。利用南瓜素：DFQL-1T复合物，我们生成了用于检测活的哺乳动物细胞和小鼠中非编码RNA和小分子靶标的基于RNA的传感器。这些研究揭示了一种荧光RNA：荧光团复合物，它可以很容易地开发成用于体内成像和传感的近红外荧光RNA标签。
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