Kentarou Sakamoto*, Yuka Yamamoto, Hiroshi Inaba and Kazunori Matsuura*,
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引用次数: 0
摘要
在透射电子显微镜(TEM)观察下,可以看到天然病毒衣壳在细胞内的自组装。通过模仿天然病毒衣壳的自组装,人们开发出了各种基于蛋白质和肽的人工纳米笼;然而,很少有研究报道这种纳米笼的细胞内自组装。我们的研究小组开发了一种β-蒽多肽,它可以在体外形成一种称为人工病毒囊的纳米笼,但这种病毒囊在细胞内的自组装尚未实现。在这里,我们设计了一种用荧光蛋白StayGold装饰的人造病毒衣壳,以观察细胞内的自组装。除了细胞的 TEM 观察和超分辨率显微镜之外,荧光各向异性测量和荧光共振能量转移成像也揭示了 StayGold 共轭的 β-Annulus 肽在细胞内自组装成了 StayGold 装饰的人工病毒衣壳。利用这些技术,我们实现了人工病毒衣壳的细胞内自组装。
Strategy toward In-Cell Self-Assembly of an Artificial Viral Capsid from a Fluorescent Protein-Modified β-Annulus Peptide
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.
期刊介绍:
The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism.
Topics may include, but are not limited to:
Design and optimization of genetic systems
Genetic circuit design and their principles for their organization into programs
Computational methods to aid the design of genetic systems
Experimental methods to quantify genetic parts, circuits, and metabolic fluxes
Genetic parts libraries: their creation, analysis, and ontological representation
Protein engineering including computational design
Metabolic engineering and cellular manufacturing, including biomass conversion
Natural product access, engineering, and production
Creative and innovative applications of cellular programming
Medical applications, tissue engineering, and the programming of therapeutic cells
Minimal cell design and construction
Genomics and genome replacement strategies
Viral engineering
Automated and robotic assembly platforms for synthetic biology
DNA synthesis methodologies
Metagenomics and synthetic metagenomic analysis
Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction
Gene optimization
Methods for genome-scale measurements of transcription and metabolomics
Systems biology and methods to integrate multiple data sources
in vitro and cell-free synthetic biology and molecular programming
Nucleic acid engineering.