Medicum, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
Department of Biomedical Engineering, School of Engineering, Duke University, Durham, NC, USA.
Nat Methods. 2023 Jan;20(1):70-74. doi: 10.1038/s41592-022-01683-0. Epub 2022 Dec 1.
Applying rational design, we developed 17 kDa cyanobacteriochrome-based near-infrared (NIR-I) fluorescent protein, miRFP718nano. miRFP718nano efficiently binds endogenous biliverdin chromophore and brightly fluoresces in mammalian cells and tissues. miRFP718nano has maximal emission at 718 nm and an emission tail in the short-wave infrared (SWIR) region, allowing deep-penetrating off-peak fluorescence imaging in vivo. The miRFP718nano structure reveals the molecular basis of its red shift. We demonstrate superiority of miRFP718nano-enabled SWIR imaging over NIR-I imaging of microbes in the mouse digestive tract, mammalian cells injected into the mouse mammary gland and NF-kB activity in a mouse model of liver inflammation.
通过合理设计,我们开发了 17 kDa 基于蓝藻视蛋白的近红外(NIR-I)荧光蛋白 miRFP718nano。miRFP718nano 能够有效地结合内源性胆绿素发色团,并在哺乳动物细胞和组织中发出明亮的荧光。miRFP718nano 的最大发射波长在 718nm 处,在短波红外(SWIR)区域有一个发射尾巴,允许在体内进行深层穿透的非峰值荧光成像。miRFP718nano 的结构揭示了其红移的分子基础。我们证明了 miRFP718nano 实现的 SWIR 成像优于微生物在小鼠消化道、注射到小鼠乳腺的哺乳动物细胞以及小鼠肝脏炎症模型中 NF-kB 活性的 NIR-I 成像。