title: Nonendocytic delivery of lipoplex nanoparticles into living cells using nanochannel electroporation.
abstract: The delivery of biomolecules, including siRNAs (≈21 bp) and large plasmids (≈10 kbp), into living cells holds a great promise for therapeutic and research applications. Lipoplex nanoparticles are popular nanocarriers for gene delivery. In conventional transfection methods, the cellular uptake of lipoplex nanoparticels occurs through the endocytosis process. The entrapment of lipoplex nanoparticles into endocytic vesicle is a major barrier in achieving efficient gene silencing and expression. Here, a novel nanochannel electroporation (NEP) method is employed to facilitate the cellular uptake and release of siRNAs/DNAs from lipoplexes. First, it is demonstrated that in a NEP device, lipoplex nanoparticles can be injected directly into the cell cytoplasm within several seconds. Specifically, it is found that lipoplexes containing MCL-1 siRNA delivered by NEP can more efficiently down-regulate the expression of MCL-1 mRNA in A549 cancer cells than conventional transfection. Quantum dot-mediated Förster resonance energy transfer (QD-FRET) reveals that lipoplexes delivered via NEP can directly release siRNA in the cytoplasm without going through the endocytosis route, which unravels the responsible mechanism for efficient gene delivery. Furthermore, the advantage of combining NEP with lipoplex nanoparticles by the successful delivery of large plasmids (pCAG2LMKOSimO, 13 kbp) into CHO cells is demonstrated.
authors.affiliation: Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric, Biomedical Devices, The Ohio State University, 174 West 18th Avenue, Columbus, OH, 43210, USA; Department of Chemical Engineering, Delft University of Technology, Julianalaan 136, 2628, BL, Delft, The Netherlands.
authors.forename: Pouyan E
authors.initials: PE
authors.lastname: Boukany
authors.forename: Yun
authors.initials: Y
authors.lastname: Wu
authors.forename: Xi
authors.initials: X
authors.lastname: Zhao
authors.forename: Kwang J
authors.initials: KJ
authors.lastname: Kwak
authors.forename: Piotr J
authors.initials: PJ
authors.lastname: Glazer
authors.forename: Kam
authors.initials: K
authors.lastname: Leong
authors.forename: L James
authors.initials: LJ
authors.lastname: Lee
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D018274:Electroporation / Q000379:methods*; D004705:Endocytosis / Q000502:physiology*; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry* / Q000493:pharmacokinetics; D015336:Molecular Probe Techniques; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry* / Q000378:metabolism; D036103:Nanotechnology / Q000379:methods; D010957:Plasmids / Q000008:administration & dosage / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics / Q000493:pharmacokinetics; D014162:Transfection / Q000379:methods*
chemical_list: D008081:Liposomes; D034741:RNA, Small Interfering
keywords: biomedical devices; electroporation; gene delivery; lipoplexes; nanofluidics