Paramyxovirus-Like Particles as Protein Delivery Vehicles.
Paramyxovirus-Like Particles as Protein Delivery Vehicles.
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作为蛋白质递送载体的副粘病毒样颗粒。
DOI：
10.1128/jvi.01030-21
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发表时间：
2021
影响因子：
5.4
通讯作者：
Schmitt,AnthonyP
中科院分区：
文献类型：
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作者：
Panthi,Santosh;Schmitt,PhuongTieu;Lorenz,FJeffrey;Stanfield,BrentA;Schmitt,AnthonyP
我们已经开发了一种灵活的平台，可以使用副粘病毒样颗粒将蛋白质输送到靶细胞内部。关键的使能特征是一个15到30个氨基酸残基的附属物，它被添加到货物蛋白中，并在病毒样颗粒(VLP)组装过程中诱导它们与病毒基质(M)蛋白结合。然后，当VLP发芽时，货物被结合到VLP中，使用通常指导病毒基因组包装的相同相互作用。该附件还可以用作表位标签，用于使用核衣壳(NP)蛋白特异性单抗进行货物检测。利用这种方法，我们产生了负载肾素酶的VLP、负载绿色荧光蛋白的VLP、负载超氧化物歧化酶的VLP和负载Cre重组酶的VLP。在每种情况下，VLP都可以有效地将其功能货物运送到靶细胞，在Cre重组酶的情况下，可以有效地运送到靶细胞核。该战略采用了两种不同的VLP生产平台，一个基于副流感病毒5(PIV5)，另一个基于Nipah病毒，在这两种情况下都可以实现高效的货物包装和交付。这些发现为开发副粘病毒样颗粒作为安全高效地将治疗性蛋白质输送到细胞和组织的工具奠定了基础。重要的是，包括转录因子和基因组编辑在内的治疗性蛋白质具有巨大的临床潜力，但目前受到限制，部分原因是安全和有效地将这些蛋白质输送到靶细胞内部的挑战。在这里，我们开发了一种基于对副粘病毒基因组包装相互作用的操纵来传递蛋白质的新策略。
We have developed a flexible platform for delivery of proteins to target cell interiors using paramyxovirus-like particles. The key enabling feature is an appendage, 15 to 30 amino acid residues in length, that is added to cargo proteins and that induces them to bind to the viral matrix (M) protein during virus-like particle (VLP) assembly. The cargo is then incorporated within the VLPs as they bud, using the same interactions that normally direct viral genome packaging. The appendage can also serve as an epitope tag for cargo detection using a nucleocapsid (NP) protein-specific monoclonal antibody. Using this approach, we generatedRenillaluciferase-loaded VLPs, green fluorescent protein-loaded VLPs, superoxide dismutase-loaded VLPs, and Cre recombinase-loaded VLPs. In each case, the VLPs could efficiently deliver their functional cargos to target cells and, in the case of Cre recombinase, to target cell nuclei. The strategy was employed using two different VLP production platforms, one based on parainfluenza virus 5 (PIV5) and the other based on Nipah virus, and in both cases efficient cargo packaging and delivery could be achieved. These findings provide a foundation for development of paramyxovirus-like particles as tools for safe and efficient delivery of therapeutic proteins to cells and tissues.IMPORTANCETherapeutic proteins including transcription factors and genome editors have enormous clinical potential but are currently limited in part due to the challenges of safely and efficiently delivering these proteins to the interiors of target cells. Here, we have developed a new strategy for protein delivery based on manipulation of paramyxovirus genome packaging interactions.