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  • Title: Stable gene expression with VSV-G pseudotyped-retrovirus vector in the rat liver.
    Author: Shiraishi M, Tomori H, Nagahama M, Taira K, Nozato E, Sugawa H, Ishida A, Muto Y.
    Journal: J Surg Res; 1999 Jun 15; 84(2):168-73. PubMed ID: 10357915.
    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. MATERIALS AND 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. CONCLUSION: 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.
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