title: Encapsulation and delivery of plasmid DNA by virus-like nanoparticles engineered from Macrobrachium rosenbergii nodavirus.
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.
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Road, Phyathai, Bangkok 10400, Thailand; School of Medicine, Walailak University, Thasala District, Nakhonsrithammarat, Thailand.
authors.forename: Pitchanee
authors.initials: P
authors.lastname: Jariyapong
authors.affiliation: Nakhonsawan Campus, Mahidol University, Nakhonsawan, Thailand. Electronic address: charoonroj.cho@mahidol.ac.th.
authors.forename: Charoonroj
authors.initials: C
authors.lastname: Chotwiwatthanakun
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Road, Phyathai, Bangkok 10400, Thailand.
authors.forename: Monsicha
authors.initials: M
authors.lastname: Somrit
authors.affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Rama 6 Road, Phyathai, Bangkok 10400, Thailand.
authors.forename: Sarawut
authors.initials: S
authors.lastname: Jitrapakdee
authors.affiliation: Department of Molecular and Cell Biology, University of California, Davis, CA 95616, United States.
authors.forename: Li
authors.initials: L
authors.lastname: Xing
authors.affiliation: Department of Molecular and Cell Biology, University of California, Davis, CA 95616, United States.
authors.forename: Holland R
authors.initials: HR
authors.lastname: Cheng
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Road, Phyathai, Bangkok 10400, Thailand. Electronic address: wattana.wee@mahidol.ac.th.
authors.forename: Wattana
authors.initials: W
authors.lastname: Weerachatyanukul
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018014:Gene Transfer Techniques / Q000295:instrumentation; D053758:Nanoparticles / Q000737:chemistry / Q000378:metabolism; D030381:Nodaviridae / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: D036022:Capsid Proteins
keywords: Biological container; Encapsulation; Macrobrachium rosenbergii nodavirus; Virus-like particle