title: A novel gene therapy to the graft organ by a rapid injection of naked DNA I: long-lasting gene expression in a rat model of limb transplantation.
abstract: BACKGROUND
  It is important to develop a nontoxic gene transfer method for immunosuppressed patients. In this study, the authors applied a nonviral gene transfer method using rapid injection of naked DNA into the graft limb in rats.
  
  
  METHODS
  Naked DNA (beta-galactosidase, luciferase, or green fluorescent protein expressing plasmid) was used to test an intravascular gene transfer approach in various conditions on the Lewis rat limb. Then, in a rat limb transplantation model, these marker genes were administered preoperatively (day -2) or perioperatively (day 0) to the graft limb by the authors' "venous protocol." The expression level of luciferase was observed over a long period using a noninvasive living image acquisition IVIS system.
  
  
  RESULTS
  Effective intravascular delivery of gene to the rat limb was achieved by a rapid bolus injection of naked DNA through the femoral caudal epigastric vein. Using this procedure, the limb graft with the marker gene perioperatively in place was safely transplanted. After limb transplantation, sustained marker gene expression was observed for more than 2 months.
  
  
  CONCLUSIONS
  This is the first report showing that the method of rapid injection of naked DNA into the limb can be applied to gene modification for organ transplantation.
authors: Division of Organ Replacement Research, Center for Molecular Medicine, Jichi Medical School, Yakushiji, Minamikawachi, Tochigi, Japan.
authors: Yuki
authors: Y
authors: Sato
authors: Takashi
authors: T
authors: Ajiki
authors: Seiichiro
authors: S
authors: Inoue
authors: Yoji
authors: Y
authors: Hakamata
authors: Takashi
authors: T
authors: Murakami
authors: Takashi
authors: T
authors: Kaneko
authors: Masafumi
authors: M
authors: Takahashi
authors: Eiji
authors: E
authors: Kobayashi
mesh_terms: D000818:Animals; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D006085:Graft Survival / Q000502:physiology*; D049452:Green Fluorescent Proteins; D006614:Hindlimb / Q000637:transplantation*; D007275:Injections, Intravenous; D008164:Luminescent Proteins / Q000235:genetics; D051381:Rats; D011917:Rats, Inbred Lew; D016038:Skin Transplantation; D013997:Time Factors; D014185:Transplantation, Isogeneic / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D001616:beta-Galactosidase