title: Stable gene expression with VSV-G pseudotyped-retrovirus vector in the rat liver.
abstract: BACKGROUND
  Pseudotyped-retrovirus-mediated gene transfer to the regenerating rat liver was investigated in vivo and the findings were compared with those for retrovirus-mediated gene transfer.
  
  
  METHODS
  Four weeks prior to gene transfer, the spleen was transpositioned to the left subcutaneous position to develop a port-splenic shunt. Twenty-four hours after a partial hepatectomy (68%) was performed, the liver was perfused in situ and kept in contact with either a pseudotyped-retrovirus vector encoding LacZ (7 x 10(7) cfu/ml, Group 1) or a retrovirus vector encoding LacZ (1 x 10(4) cfu/ml, Group 2) for 30 min. The animals were sacrificed at various points after gene transfer, and X-gal staining, reversed polymerase chain reaction (RT-PCR), and ONPG assay were performed to detect the transferred LacZ cDNA.
  
  
  RESULTS
  In X-gal staining, the transferred LacZ cDNA started to show a strong beta-galactosidase activity in 30 to 50% of the hepatocytes at 3 days after gene transfer. Positive staining continued to be recognized until 28 days with a slight decrease in its intensity thereafter. On the other hand, Group 2 animals showed weak staining, which was observed in about 10 to 15% of the hepatocytes from 3 days after gene transfer and then decreased thereafter. In RT-PCR, positive mRNA of LacZ was detected constitutively until 28 days after gene transfer in Group 1, whereas two-thirds of the samples showed a negative band in Groups 2 at 3 days after gene transfer.
  
  
  CONCLUSIONS
  In conclusion, the pseudotyped-retrovirus vector was useful in establishing a stable and strong expression of the in vivo gene transfer, while targeting the regenerating liver.
authors.affiliation: First Department of Surgery, University of the Ryukyus, Okinawa, 903-0215, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Shiraishi
authors.forename: H
authors.initials: H
authors.lastname: Tomori
authors.forename: M
authors.initials: M
authors.lastname: Nagahama
authors.forename: K
authors.initials: K
authors.lastname: Taira
authors.forename: E
authors.initials: E
authors.lastname: Nozato
authors.forename: H
authors.initials: H
authors.lastname: Sugawa
authors.forename: A
authors.initials: A
authors.lastname: Ishida
authors.forename: Y
authors.initials: Y
authors.lastname: Muto
mesh_terms: D000818:Animals; D002863:Chromogenic Compounds; D005697:Galactosides; D015870:Gene Expression / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics*; D007211:Indoles; D007763:Lac Operon / Q000235:genetics; D008099:Liver / Q000378:metabolism / Q000502:physiology*; D008562:Membrane Glycoproteins; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012190:Retroviridae / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013194:Staining and Labeling; D014759:Viral Envelope Proteins / Q000235:genetics*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D002863:Chromogenic Compounds; C028391:G protein, vesicular stomatitis virus; D005697:Galactosides; D007211:Indoles; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside