title: Anionic/Zwitterionic Lipid-Based Gene Vectors of pDNA.
abstract: The use of anionic lipids (ALs) as non-viral gene vectors depicts a promising alternative to cationic lipids (CLs) since they are more biocompatible and present lower levels of phagocytosis by macrophages. Several experimental methods, such as electrophoretic mobility (ζ-potential), gel electrophoresis, small-angle X-ray scattering (SAXS), fluorescence and confocal fluorescence microscopies (FM and CFM), flow assisted cell sorting-flow cytometry (FACS-FCM), and cell viability/cytotoxicity assays can be used for a complete physicochemical and biochemical characterization of lipoplexes formed by an AL, a zwitterionic lipid (ZL), and a plasmid DNA (pDNA), their electrostatic interaction being necessarily mediated by divalent cations, such as Ca(2+). In the present chapter, we summarize the protocols optimized for the mentioned characterization techniques.
authors.affiliation: Grupo de Química Coloidal y Supramolecular, Departamento de Química Física I, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Ciudad Universitaris s/n, 28040, Madrid, Spain.
authors.forename: Ana L
authors.initials: AL
authors.lastname: Barrán-Berdón
authors.affiliation: Grupo de Química Coloidal y Supramolecular, Departamento de Química Física I, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Ciudad Universitaris s/n, 28040, Madrid, Spain.
authors.forename: Emilio
authors.initials: E
authors.lastname: Aicart
authors.affiliation: Grupo de Química Coloidal y Supramolecular, Departamento de Química Física I, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Ciudad Universitaris s/n, 28040, Madrid, Spain. junquera@quim.ucm.es.
authors.forename: Elena
authors.initials: E
authors.lastname: Junquera
mesh_terms: D000838:Anions / Q000737:chemistry*; D002118:Calcium; D002469:Cell Separation; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D005434:Flow Cytometry; D015316:Genetic Therapy; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D053838:Scattering, Small Angle; D014162:Transfection; D014961:X-Ray Diffraction
chemical_list: D000838:Anions; D008055:Lipids; D008081:Liposomes; D004247:DNA; D002118:Calcium
keywords: Anionic lipids; Flow cytometry; Fluorescence microscopy; Gel electrophoresis; Lipoplexes; Liposomes; Non-viral gene vectors; SAXS; pDNA; ζ-potential