Retroviral-mediated gene transfer into hepatocytes in vivo.
Stable gene transfer into hepatocytes might be used to compensate for a genetic deficiency affecting liver function or to deliver diffusible factors into the blood stream. In rats, we have combined retroviral-mediated gene transfer with a surgical procedure in which the liver is temporarily excluded from the circulation and infected in vivo. Partial hepatectomy was performed 24-48 hr before perfusion with virus to induce hepatocyte division and facilitate viral integration. A helper-free recombinant retrovirus coding for beta-galactosidase with nuclear localization was used to score cells that expressed the transgene. For at least 3 months after gene transfer, up to 5% of hepatocytes expressed nuclear beta-galactosidase. Whereas in vitro reimplantation of genetically modified hepatocytes has proved to be inefficient in stably transferring genes into the liver, our approach provides a feasible alternative.
Ferry
N
N
Laboratoire Rétrovirus et Transfert Génétique, Institut Pasteur, Paris, France.
Duplessis
O
O
Houssin
D
D
Danos
O
O
Heard
J M
JM
D000818:Animals; D001483:Base Sequence; D002451:Cell Compartmentation; D015870:Gene Expression; D005822:Genetic Vectors; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008099:Liver / Q000502:physiology*; D008115:Liver Regeneration; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D010477:Perfusion; D016133:Polymerase Chain Reaction; D051381:Rats; D011917:Rats, Inbred Lew; D011993:Recombinant Fusion Proteins; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase