Li Chenge, Plamont Marie-Aude, Sladitschek Hanna L, Rodrigues Vanessa, Aujard Isabelle, Neveu Pierre, Le Saux Thomas, Jullien Ludovic, Gautier Arnaud
École Normale Supérieure , PSL Research University , UPMC Univ Paris 06 , CNRS , Département de Chimie , PASTEUR , 24 rue Lhomond , 75005 Paris , France.
Sorbonne Universités , UPMC Univ Paris 06 , ENS , CNRS , PASTEUR , 75005 Paris , France . Email:
Chem Sci. 2017 Aug 1;8(8):5598-5605. doi: 10.1039/c7sc01364g. Epub 2017 May 30.
Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST, hereafter called FAST) is a 14 kDa protein tag giving a bright green-yellow fluorescent complex upon interaction with the fluorogenic dye 4-hydroxy-3-methylbenzylidene rhodanine (HMBR). Here, we report a collection of fluorogens enabling tuning of the fluorescence color of FAST from green-yellow to orange and red. Beyond allowing the multicolor imaging of FAST-tagged proteins in live cells, these fluorogens enable dynamic color switching because of FAST's reversible labeling. This unprecedented behavior allows for selective detection of FAST-tagged proteins in cells expressing both green and red fluorescent species through two-color cross-correlation, opening up exciting prospects to overcome spectral crowding and push the frontiers of multiplexed imaging.
黄色荧光激活与吸收转移标签（Y-FAST，以下简称FAST）是一种14 kDa的蛋白质标签，与荧光染料4-羟基-3-甲基苄叉罗丹宁（HMBR）相互作用时会形成亮绿黄色荧光复合物。在此，我们报告了一系列荧光原，它们能够将FAST的荧光颜色从绿黄色调谐为橙色和红色。这些荧光原不仅能实现活细胞中FAST标记蛋白的多色成像，还因FAST的可逆标记实现动态颜色切换。这种前所未有的特性使得通过双色互相关在同时表达绿色和红色荧光物质的细胞中选择性检测FAST标记蛋白成为可能，为克服光谱拥挤和推动多重成像前沿开辟了令人兴奋的前景。
