Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, 100101, P. R. China.
Department of Pharmacology, Weill Cornell Medicine, Cornell University, New York, NY, 10065, USA.
Angew Chem Int Ed Engl. 2021 Nov 2;60(45):24153-24161. doi: 10.1002/anie.202108338. Epub 2021 Oct 4.
Fluorogenic aptamers can potentially show minimal photobleaching during continuous irradiation since any photobleached fluorophore can exchange with fluorescent dyes in the media. However, fluorophores have not been designed to maximize "fluorophore recycling." Here we describe TBI, a novel fluorophore for the Broccoli fluorogenic aptamer. Previous fluorophores either fail to rapidly dissociate when they undergo photobleaching via cis-trans isomerization, or bind slowly, resulting in extended periods after dissociation of the photobleached fluorophore when no fluorophore is bound. By contrast, photobleached TBI dissociates rapidly from Broccoli, and TBI from the media rapidly replaces dissociated photobleached fluorophore. Using TBI, Broccoli exhibits markedly enhanced fluorescence in cells during continuous imaging. These data show that designing fluorophores to optimize fluorophore recycling can lead to enhanced fluorescence of fluorogenic aptamers.
荧光适体在连续照射下可能表现出最小的光漂白，因为任何光漂白的荧光团都可以与介质中的荧光染料交换。然而，荧光团的设计并没有最大限度地提高“荧光团的回收利用”。在这里，我们描述了 TBI，这是一种用于西兰花荧光适体的新型荧光团。以前的荧光团要么在顺反异构导致光漂白时不能快速解离，要么结合缓慢，导致光漂白的荧光团解离后，在没有荧光团结合的情况下，会有很长一段时间。相比之下，光漂白的 TBI 从西兰花中迅速解离，而来自介质的 TBI 迅速取代解离的光漂白荧光团。使用 TBI，西兰花在连续成像过程中在细胞内表现出明显增强的荧光。这些数据表明，设计荧光团以优化荧光团的回收利用可以提高荧光适体的荧光强度。
