Medicum, Faculty of Medicine, University of Helsinki, Helsinki, 00290, Finland.
Present address: Institute of Molecular and Cellular Biology, Novosibirsk, 630090, Russia.
Chembiochem. 2018 Jun 18;19(12):1334-1340. doi: 10.1002/cbic.201700642. Epub 2018 Apr 14.
Near-infrared (NIR) light-inducible binding of bacterial phytochrome BphP1 to its engineered partner, QPAS1, is used for optical protein regulation in mammalian cells. However, there are no data on the application of the BphP1-QPAS1 pair in cells derived from various mammalian tissues. Here, we tested the functionality of two BphP1-QPAS1-based optogenetic tools-an NIR- and blue-light-sensing system for control of protein localization (iRIS) and an NIR light-sensing system for transcription activation (TA)-in several cell types, including cortical neurons. We found that the performance of these optogenetic tools often relied on physiological properties of a specific cell type, such as nuclear transport, which could limit the applicability of the blue-light-sensitive component of iRIS. In contrast, the NIR-light-sensing component of iRIS performed well in all tested cell types. The TA system showed the best performance in cervical cancer (HeLa), bone cancer (U-2 OS), and human embryonic kidney (HEK-293) cells. The small size of the QPAS1 component allowed the design of adeno-associated virus (AAV) particles, which were applied to deliver the TA system to neurons.
近红外（NIR）光诱导的细菌叶绿素 BphP1 与其工程化伙伴 QPAS1 的结合被用于哺乳动物细胞中的光学蛋白质调控。然而，关于 BphP1-QPAS1 对在各种哺乳动物组织来源的细胞中的应用，尚无数据。在这里，我们测试了基于 BphP1-QPAS1 的两种光遗传学工具的功能——一种用于控制蛋白质定位的近红外和蓝光感应系统（iRIS）和一种用于转录激活的近红外光感应系统（TA）——在几种细胞类型中，包括皮质神经元。我们发现，这些光遗传学工具的性能通常依赖于特定细胞类型的生理特性，例如核转运，这可能限制了 iRIS 中蓝光敏感组件的适用性。相比之下，iRIS 的近红外光感应组件在所有测试的细胞类型中表现良好。TA 系统在宫颈癌（HeLa）、骨癌（U-2 OS）和人胚肾（HEK-293）细胞中表现最佳。QPAS1 组件的体积小，允许设计腺相关病毒（AAV）颗粒，该颗粒被用于将 TA 系统递送至神经元。
