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罗氏沼虾诺达病毒样纳米颗粒的封装与质粒 DNA 的递送。

Encapsulation and delivery of plasmid DNA by virus-like nanoparticles engineered from Macrobrachium rosenbergii nodavirus.

机构信息

Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Road, Phyathai, Bangkok 10400, Thailand; School of Medicine, Walailak University, Thasala District, Nakhonsrithammarat, Thailand.

Nakhonsawan Campus, Mahidol University, Nakhonsawan, Thailand.

出版信息

Virus Res. 2014 Jan 22;179:140-6. doi: 10.1016/j.virusres.2013.10.021. Epub 2013 Oct 31.

Abstract

Virus-like particles (VLPs) are potential candidates in developing biological containers for packaging therapeutic or biologically active agents. Here, we expressed Macrobrachium rosenbergii nodavirus (MrNv) capsid protein (encoding amino acids M1-N371 with 6 histidine residuals) in an Escherichia coli BL21(DE3). These easily purified capsid protein self-assembled into VLPs, and disassembly/reassembly could be controlled in a calcium-dependent manner. Physically, MrNv VLPs resisted to digestive enzymes, a property that should be advantageous for protection of active compounds against harsh conditions. We also proved that MrNv VLPs were capable of encapsulating plasmid DNA in the range of 0.035-0.042 mol ratio (DNA/protein) or 2-3 plasmids/VLP (assuming that MrNV VLPs is T=1, i made up of 60 capsid monomers). These VLPs interacted with cultured insect cells and delivered loaded plasmid DNA into the cells as shown by green fluorescent protein (GFP) reporter. With many advantageous properties including self-encapsulation, MrNv VLPs are good candidates for delivery of therapeutic agents.

摘要

病毒样颗粒(VLPs)是一种有潜力的生物容器,可用于包装治疗性或具有生物活性的药物。在这里,我们在大肠杆菌 BL21(DE3)中表达了罗氏沼虾诺达病毒(MrNv)衣壳蛋白(编码 M1-N371 氨基酸,带有 6 个组氨酸残基)。这些衣壳蛋白可以自行组装成 VLPs,并且可以通过钙离子依赖的方式控制其解组装/组装。从物理角度来看,MrNv VLPs 能够抵抗消化酶,这一特性有利于保护活性化合物免受恶劣条件的影响。我们还证明,MrNv VLPs 能够在 0.035-0.042 摩尔比(DNA/蛋白)或 2-3 个质粒/VLP 的范围内封装质粒 DNA(假设 MrNV VLPs 是 T=1,由 60 个衣壳单体组成)。这些 VLPs 与培养的昆虫细胞相互作用,并通过绿色荧光蛋白(GFP)报告基因将负载的质粒 DNA 递送到细胞中。MrNv VLPs 具有自包封等许多优点,是治疗药物递送的良好候选物。

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