<br>明亮稳定的青色荧光 RNA 可在活大肠杆菌中实现多色 RNA 成像,
明亮稳定的青色荧光 RNA 可在活大肠杆菌中实现多色 RNA 成像
Small
(
IF
13.0
)
Pub Date : 2024-10-28
, DOI:
10.1002/smll.202405165
Li Jiang, Fangting Zuo, Yuanyuan Pan, Ruilong Li, Yajie Shi, Xinyi Huang, Dasheng Zhang, Yingping Zhuang, Yuzheng Zhao, Qiuning Lin, Yi Yang, Linyong Zhu, Xianjun Chen
荧光 RNA （FR） 是结合并激活其同源小荧光染料的 RNA 适体，石材翻新13825404095为活细胞中的 RNA 成像提供了一种特别有用的方法。尽管 FR 的调色板大大扩展，但与目前可用的 FR 具有良好的生物相容性和双正交性的明亮稳定的青色 FR 仍然是可取的，但尚未开发。本文描述了 Myosotis 的发育，一种 RNA 适体，在结合一种称为 DBT 的新型 GFP 发色团样荧光团时发出明亮的青色荧光。肌巢菌对 DBT 具有纳摩尔亲和力，并且对镁的折叠依赖性较弱。进一步的研究表明，Myosotis-DBT 复合物具有较长的荧光寿命、良好的光稳定性，并增强了细胞亮度。进一步表明，Myosotis-DBT 与 Pepper 和 Clivia FR 双正交，允许对活细菌中的 RNA 进行多重荧光成像。肌层还可用于对活细菌中的 mRNA 进行成像，揭示 mRNA 翻译与稳定性之间的潜在耦合。据信，这种青色 FR 将成为研究不同生物过程背后的 RNA 功能和机制的有用工具。
"点击查看英文标题和摘要"
Bright and Stable Cyan Fluorescent RNA Enables Multicolor RNA Imaging in Live Escherichia coli
Fluorescent RNAs (FRs), which are RNA aptamers that bind and activate their cognate small fluorogenic dyes, have provided a particularly useful approach for imaging RNAs in live cells. Although the color palette of FRs is greatly expanded, a bright and stable cyan FR with good biocompatibility and biorthogonality with currently available FRs remains desirable but is not yet developed. Herein, the development of Myosotis is described, an RNA aptamer that emits bright cyan fluorescence upon binding a novel GFP chromophore‐like fluorophore called DBT. Myosotis has a nanomolar affinity for DBT and shows a weak dependence on magnesium for folding. Further studies reveal that the Myosotis‐DBT complex has a long fluorescence lifetime, good photostability, and enhance cellular brightness. It is further shown that Myosotis‐DBT is biorthogonal to Pepper and Clivia FRs, allowing multiplex fluorescence imaging of RNA in live bacteria. Myosotis can also use to image mRNA in live bacteria, revealing potential coupling between mRNA translation and stability. It is believed that this cyan FR will be a useful tool for studying the functionality and mechanism of RNA underlying diverse biological processes.