Department of Chemistry and Biotechnology, Graduate School of Engineering, Tottori University, Tottori 680-8552, Japan.
Centre for Research on Green Sustainable Chemistry, Tottori University, Tottori 680-8552, Japan.
ACS Synth Biol. 2024 Jun 21;13(6):1842-1850. doi: 10.1021/acssynbio.4c00135. Epub 2024 May 10.
In-cell self-assembly of natural viral capsids is an event that can be visualized under transmission electron microscopy (TEM) observations. By mimicking the self-assembly of natural viral capsids, various artificial protein- and peptide-based nanocages were developed; however, few studies have reported the in-cell self-assembly of such nanocages. Our group developed a β-Annulus peptide that can form a nanocage called artificial viral capsid in vitro, but in-cell self-assembly of the capsid has not been achieved. Here, we designed an artificial viral capsid decorated with a fluorescent protein, StayGold, to visualize in-cell self-assembly. Fluorescence anisotropy measurements and fluorescence resonance energy transfer imaging, in addition to TEM observations of the cells and super-resolution microscopy, revealed that StayGold-conjugated β-Annulus peptides self-assembled into the StayGold-decorated artificial viral capsid in a cell. Using these techniques, we achieved the in-cell self-assembly of an artificial viral capsid.
细胞内自组装天然病毒衣壳是一种可以在透射电子显微镜（TEM）观察下可视化的事件。通过模拟天然病毒衣壳的自组装，开发了各种基于人工蛋白质和肽的纳米笼；然而，很少有研究报道过这种纳米笼的细胞内自组装。我们小组开发了一种β-Annulus 肽，它可以在体外形成一种称为人工病毒衣壳的纳米笼，但尚未实现衣壳的细胞内自组装。在这里，我们设计了一种带有荧光蛋白 StayGold 的人工病毒衣壳，以可视化细胞内自组装。荧光各向异性测量和荧光共振能量转移成像，以及细胞的 TEM 观察和超分辨率显微镜，表明 StayGold 缀合的β-Annulus 肽在细胞内自组装成带有 StayGold 的人工病毒衣壳。使用这些技术，我们实现了人工病毒衣壳的细胞内自组装。
