A genetically targetable near-infrared photosensitizer.
A genetically targetable near-infrared photosensitizer.
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DOI：
10.1038/nmeth.3735
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发表时间：
2016-03
期刊：
影响因子：
48
通讯作者：
Bruchez MP
中科院分区：
文献类型：
--
作者：
He J;Wang Y;Missinato MA;Onuoha E;Perkins LA;Watkins SC;St Croix CM;Tsang M;Bruchez MP
在光照下，光敏剂分子产生活性氧物种(ROS)，可用于活细胞的功能操作，包括蛋白质失活、靶向损伤引入和细胞消融。到目前为止，使用的光敏剂要么是外源的，导致递送和移除挑战，要么是基因编码的蛋白质形成或结合天然的光敏分子，导致细胞内具有结构性活性的光敏剂。通过将重原子取代的荧光染料与基因编码的氟原激活蛋白(FAP)结合，我们展示了一种“按需”激活的光敏剂，仅当FAP结合并用近红外光激活时才产生单线态氧和荧光。这种靶向和激活的光敏剂(TAPS)方法能够使培养细胞中的蛋白质失活和靶向细胞杀伤，并在活的幼体和成年斑马鱼中快速靶向消融。这种FAP-TAPS光敏剂模块的近红外激发和发射为光敏剂蛋白质提供了一个新的光谱范围，对于成像、操作和生物体内深处的细胞消融非常有用。
Upon illumination, photosensitizer molecules produce reactive oxygen species (ROS) that can be utilized for functional manipulation of living cells, including protein inactivation, targeted damage introduction, and cellular ablation. Photosensitizers used to date have been either exogenous, resulting in delivery and removal challenges, or genetically encoded proteins that form or bind a native photosensitizing molecule, resulting in a constitutively active photosensitizer inside the cell. By binding a heavy-atom substituted fluorogenic dye with a genetically encoded Fluorogen Activating Protein (FAP), we demonstrate an ‘on-demand’ activated photosensitizer that produces singlet oxygen and fluorescence only when FAP-bound and activated with near infrared light. This Targeted and Activated Photosensitizer (TAPs) approach enables protein inactivation and targeted cell killing in cultured cells and rapid targeted lineage ablation in living larval and adult zebrafish. The near-infrared excitation and emission of this FAP-TAPs photosensitizer module provides a new spectral range for photosensitizer proteins, useful for imaging, manipulation and cellular ablation deep within living organisms.