title: Development of high-concentration lipoplexes for in vivo gene function studies in vertebrate embryos.
abstract: Here we report that highly concentrated cationic lipid/helper lipid-nucleic acid complexes (lipoplexes) can facilitate reproducible delivery of a variety of oligonucleotides and plasmids to chicken embryos or to mouse embryonic mesenchyme. Specifically, liposomes composed of N,N-dioleyl-N,N-dimethylammonium chloride (DODAC)/1,2 dioleoyl glycero-3-phosphorylethanolamine (DOPE) prepared at 18-mM concentrations produced high levels of transfection of exogenous genes in vivo and in vitro. Furthermore, we report sufficient uptake of plasmids expressing interference RNA to decrease expression of both exogenous and endogenous genes. The simplicity of preparation, implementation, and relatively low toxicity of this transfection reagent make it an attractive alternative for developmental studies in post-gastrulation vertebrate embryos.
authors.affiliation: Department of Oral Health Sciences, Life Sciences Centre, The University of British Columbia, Vancouver Canada.
authors.forename: Poongodi
authors.initials: P
authors.lastname: Geetha-Loganathan
authors.forename: Suresh
authors.initials: S
authors.lastname: Nimmagadda
authors.forename: Ismail
authors.initials: I
authors.lastname: Hafez
authors.forename: Katherine
authors.initials: K
authors.lastname: Fu
authors.forename: Pieter R
authors.initials: PR
authors.lastname: Cullis
authors.forename: Joy M
authors.initials: JM
authors.lastname: Richman
mesh_terms: D000818:Animals; D002642:Chick Embryo; D004622:Embryo, Mammalian; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry*; D008648:Mesoderm / Q000378:metabolism; D051379:Mice; D010714:Phosphatidylethanolamines / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000737:chemistry; D014162:Transfection / Q000379:methods*
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; C015831:dimethyldioctadecylammonium