Kasatkina Ludmila A, Ma Chenshuo, Sheng Huaxin, Lowerison Matthew, Menozzi Luca, Baloban Mikhail, Tang Yuqi, Xu Yirui, Humayun Lucas, Vu Tri, Song Pengfei, Yao Junjie, Verkhusha Vladislav V
bioRxiv. 2024 Oct 18:2024.10.18.619161. doi: 10.1101/2024.10.18.619161.
We developed near-infrared (NIR) photoacoustic and fluorescence probes, as well as optogenetic tools from bacteriophytochromes, and enhanced their performance using biliverdin reductase-A knock-out model (Blvra-/-). Blvra-/- elevates endogenous heme-derived biliverdin chromophore for bacteriophytochrome-derived NIR constructs. Consequently, light-controlled transcription with IsPadC-based optogenetic tool improved up to 25-fold compared to wild-type cells, with 100-fold activation in Blvra-/- neurons. , light-induced insulin production in Blvra-/- reduced blood glucose in diabetes by ∼60%, indicating high potential for optogenetic therapy. Using 3D photoacoustic, ultrasound, and two-photon fluorescence imaging, we overcame depth limitations of recording NIR probes. We achieved simultaneous photoacoustic imaging of DrBphP in neurons and super-resolution ultrasound localization microscopy of blood vessels ∼7 mm deep in the brain, with intact scalp and skull. Two-photon microscopy provided cell-level resolution of miRFP720-expressing neurons ∼2.2 mm deep. Blvra-/- significantly enhances efficacy of biliverdin-dependent NIR systems, making it promising platform for interrogation and manipulation of biological processes.
我们开发了近红外（NIR）光声和荧光探针，以及源自细菌光敏色素的光遗传学工具，并使用胆绿素还原酶-A基因敲除模型（Blvra-/-）提高了它们的性能。Blvra-/-可提高细菌光敏色素衍生的近红外构建体的内源性血红素衍生胆绿素发色团。因此，与野生型细胞相比，基于IsPadC的光遗传学工具的光控转录提高了25倍，在Blvra-/-神经元中激活了100倍。此外，Blvra-/-中光诱导的胰岛素产生使糖尿病患者的血糖降低了约60%，表明光遗传学治疗具有很高的潜力。通过三维光声、超声和双光子荧光成像，我们克服了记录近红外探针的深度限制。我们实现了对神经元中DrBphP的同步光声成像以及对大脑中约7毫米深处血管的超分辨率超声定位显微镜成像，头皮和颅骨完整。双光子显微镜提供了对约2.2毫米深处表达miRFP720的神经元的细胞水平分辨率。Blvra-/-显著提高了胆绿素依赖性近红外系统的功效，使其成为用于研究和操纵生物过程的有前景的平台。
