A bacterial phytochrome-based optogenetic system controllable with near-infrared light.
A bacterial phytochrome-based optogenetic system controllable with near-infrared light.
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DOI：
10.1038/nmeth.3864
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发表时间：
2016-07
期刊：
影响因子：
48
通讯作者：
Verkhusha VV
中科院分区：
文献类型：
--
作者：
Kaberniuk AA;Shemetov AA;Verkhusha VV
光介导的蛋白质相互作用调控代谢途径是光遗传学的重要途径。在这里，我们报道了第一个基于细菌光敏色素BphP1与其天然伴侣PpsR2之间可逆光诱导结合的光遗传系统。我们在体外和哺乳动物细胞中广泛表征了BphP1-PpsR2相互作用，然后利用它将靶蛋白转运到特定的细胞区室，如质膜和细胞核。应用这种方法，我们实现了细胞形态的光控，导致细胞面积的大幅增加。接下来，我们在小鼠的培养细胞、皮下和深层组织中分别用40倍、32倍和5.7倍的对比证明了光诱导基因的表达。BphP1-PpsR2光遗传系统的独特特征是其对740 - 780nm近红外光的敏感性，能够利用包括哺乳动物在内的真核生物的内源性胆绿素发色团，以及与蓝光光遗传系统的光谱兼容性。
Light-mediated control of protein-protein interactions to regulate metabolic pathways is an important approach of optogenetics. Here, we report the first optogenetic system based on a reversible light-induced binding between a bacterial phytochrome BphP1 and its natural partner PpsR2 from Rhodopseudomonas palustris bacteria. We extensively characterized the BphP1–PpsR2 interaction both in vitro and in mammalian cells, and then used it to translocate target proteins to specific cellular compartments, such as plasma membrane and nucleus. Applying this approach we achieved a light-control of cell morphology resulting in the substantial increase of cell area. We next demonstrated the light-induced gene expression with the 40-fold contrast in cultured cells, 32-fold subcutaneously and 5.7-fold in deep tissues in mice. The unique characteristics of the BphP1–PpsR2 optogenetic system are its sensitivity to 740–780 nm near-infrared light, ability to utilize an endogenous biliverdin chromophore in eukaryotes including mammals, and spectral compatibility with blue-light optogenetic systems.