适体
化学
生物物理学
螺旋(腹足类)
核糖核酸
生物素化
荧光团
碱基对
小分子
荧光
G-四倍体
生物
DNA
生物化学
分子生物学
基因
物理
蜗牛
量子力学
生态学
作者
Kaiyi Huang,Xianjun Chen,Chunyan Li,Qianqian Song,Huiwen Li,Linyong Zhu,Yi Yang,Aiming Ren
标识
DOI:10.1038/s41589-021-00884-6
摘要
Pepper fluorescent RNAs are a recently reported bright, stable and multicolor fluorogenic aptamer tag that enable imaging of diverse RNAs in live cells. To investigate the molecular basis of the superior properties of Pepper, we determined the structures of complexes of Pepper aptamer bound with its cognate HBC or HBC-like fluorophores at high resolution by X-ray crystallography. The Pepper aptamer folds in a monomeric non-G-quadruplex tuning-fork-like architecture composed of a helix and one protruded junction region. The near-planar fluorophore molecule intercalates in the middle of the structure and is sandwiched between one non-G-quadruplex base quadruple and one noncanonical G·U wobble helical base pair. In addition, structure-based mutational analysis is evaluated by in vitro and live-cell fluorogenic detection. Taken together, our research provides a structural basis for demystifying the fluorescence activation mechanism of Pepper aptamer and for further improvement of its future application in RNA visualization.
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