Engineered fluorogenic HaloTag ligands for turn-on labelling in live cells.
Engineered fluorogenic HaloTag ligands for turn-on labelling in live cells.
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工程化荧光 HaloTag 配体，用于活细胞中的开启标记。
DOI：
10.1039/d3cc05536a
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Kritzer,JoshuaA
中科院分区：
文献类型：
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作者：
Lampkin,BryanJ;Kritzer,JoshuaA
近年来，基于限制非辐射衰减途径的有机荧光团的设计已经有了显著的改进。我们试图将这种理解扩展到苯并噻二唑，其已被用作自标记蛋白HaloTag的开启荧光底物。当与HaloTag共轭时，苯并噻二唑驻留在一个狭窄的隧道中，排除了扭曲的内部电荷转移，这使我们能够探索对其他非辐射衰变途径的空间和电子效应。通过最小化非辐射衰减和与细胞组分的非特异性相互作用，我们产生了改进的开启染料，其荧光比细胞中的背景增加了136倍。
Recent years have seen dramatic improvements in the design of organic fluorophores based on limiting non-radiative decay pathways. We sought to extend this understanding to benzothiadiazoles that have been used as turn-on fluorescent substrates for the self-labeling protein HaloTag. When conjugated to HaloTag, the benzothiadiazoles reside in a narrow tunnel that precludes twisted internal charge transfer, which allowed us to explore steric and electronic effects on other non-radiative decay pathways. By minimizing both non-radiative decay and nonspecific interactions with cellular components, we produced improved turn-on dyes with 136-fold increase in fluorescence over background in cells.