title: Design of lipid nanoparticles for in vitro and in vivo delivery of plasmid DNA.
abstract: Lipid nanoparticles (LNPs) containing distearoylphosphatidlycholine (DSPC), and ionizable amino-lipids such as dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA) are potent siRNA delivery vehicles in vivo. Here we explore the utility of similar LNP systems as transfection reagents for plasmid DNA (pDNA). It is shown that replacement of DSPC by unsaturated PCs and DLin-MC3-DMA by the related lipid DLin-KC2-DMA resulted in highly potent transfection reagents for HeLa cells in vitro. Further, these formulations exhibited excellent transfection properties in a variety of mammalian cell lines and transfection efficiencies approaching 90% in primary cell cultures. These transfection levels were equal or greater than achieved by Lipofectamine, with much reduced toxicity. Finally, microinjection of LNP-eGFP into the limb bud of a chick embryo resulted in robust reporter-gene expression. It is concluded that LNP systems containing ionizable amino lipids can be highly effective, non-toxic pDNA delivery systems for gene expression both in vitro and in vivo.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Jayesh A
authors.initials: JA
authors.lastname: Kulkarni
authors.affiliation: Department of Oral Health Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Johnathan Layne
authors.initials: JL
authors.lastname: Myhre
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Sam
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Yuen Yi C
authors.initials: YYC
authors.lastname: Tam
authors.affiliation: Department of Oral Health Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Adrian
authors.initials: A
authors.lastname: Danescu
authors.affiliation: Department of Oral Health Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Joy M
authors.initials: JM
authors.lastname: Richman
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada. Electronic address: pieterc@mail.ubc.ca.
authors.forename: Pieter R
authors.initials: PR
authors.lastname: Cullis
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002642:Chick Embryo; D004247:DNA / Q000737:chemistry*; D016503:Drug Delivery Systems; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry*; D014162:Transfection
chemical_list: D008055:Lipids; D004247:DNA
keywords: Gene delivery; Gene expression; Lipid nanoparticles; Nanomedicine; Plasmid