title: Up-Scaled Synthesis and Characterization of Nonviral Gene Delivery Particles for Transient In Vitro and In Vivo Transgene Expression.
abstract: Polyethylenimine-based polyplexes are promising nonviral gene delivery systems for preclinical and clinical applications. Pipette-based polyplexing is associated with several disadvantages, such as batch-to-batch variability, restriction to smaller volumes, and variable gene delivery results. The present protocol describes syringe-pump-mediated upscaled synthesis of well-defined gene delivery nanoparticles capable of efficient in vitro and in vivo gene delivery. Syringe-pump-based synthesis ensures controlled mixing, upscaling, and reproducible gene delivery. Nanoparticle tracking analysis of the upscaled formulations involved single nanoparticle tracking, thereby generating highly resolved biophysical characterization. Gene delivery performance was investigated by luciferase gene expression in cells and three-dimensional bioluminescence imaging in mice.
authors.affiliation: MMCT Laboratory of Macromolecular Cancer Therapeutics, Department for Pharmaceutical Chemistry, Faculty Center for Pharmacy, University of Vienna , Vienna, Austria .
authors.forename: Alexander
authors.initials: A
authors.lastname: Taschauer
authors.affiliation: MMCT Laboratory of Macromolecular Cancer Therapeutics, Department for Pharmaceutical Chemistry, Faculty Center for Pharmacy, University of Vienna , Vienna, Austria .
authors.forename: Antonia
authors.initials: A
authors.lastname: Geyer
authors.affiliation: MMCT Laboratory of Macromolecular Cancer Therapeutics, Department for Pharmaceutical Chemistry, Faculty Center for Pharmacy, University of Vienna , Vienna, Austria .
authors.forename: Sebastian
authors.initials: S
authors.lastname: Gehrig
authors.affiliation: MMCT Laboratory of Macromolecular Cancer Therapeutics, Department for Pharmaceutical Chemistry, Faculty Center for Pharmacy, University of Vienna , Vienna, Austria .
authors.forename: Julia
authors.initials: J
authors.lastname: Maier
authors.affiliation: MMCT Laboratory of Macromolecular Cancer Therapeutics, Department for Pharmaceutical Chemistry, Faculty Center for Pharmacy, University of Vienna , Vienna, Austria .
authors.forename: Haider
authors.initials: H
authors.lastname: Sami
authors.affiliation: MMCT Laboratory of Macromolecular Cancer Therapeutics, Department for Pharmaceutical Chemistry, Faculty Center for Pharmacy, University of Vienna , Vienna, Austria .
authors.forename: Manfred
authors.initials: M
authors.lastname: Ogris
mesh_terms: D000818:Animals; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry* / Q000627:therapeutic use; D011094:Polyethyleneimine / Q000737:chemistry* / Q000627:therapeutic use; D014162:Transfection; D019076:Transgenes / Q000235:genetics
chemical_list: D011094:Polyethyleneimine; D008156:Luciferases