Recombinant avian adeno-associated virus: transgene expression in vivo and enhancement of expression in vitro.
Recombinant avian adeno-associated viruses coding for the LacZ gene were used to inoculate embryonating chicken eggs, to assess the usefulness of the system for the expression of a transgene in vivo. The results obtained indicate significantly higher levels of expression of the reporter gene at various time intervals in the embryos inoculated with the recombinant virus in comparison with the mock-inoculated controls. At the embryo level, significant differences were evident at 120 hr postinoculation; hatched chicks showed transgene expression up to 14 days of age. In a second experiment, different cell-line cultures were transfected with plasmids encoding for a reporter gene flanked by the avian adeno-associated virus inverted terminal repeats (ITR), either alone or in the presence of the major nonstructural proteins of the virus (Rep 78/68) to assess the ability of these proteins and DNA elements to enhance gene expression. Results indicate that the inclusion of the viral ITR alone or during coexpression of the Rep proteins significantly enhances the expression of the transgene in all cell lines tested, as evidenced by the detection of the beta-galacrosidase protein through chemiluminescence reactions and staining of transfected monolayers.
Estevez
Carlos
C
Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens 30602, USA.
Villegas
Pedro
P
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics* / Q000821:virology*; D000229:Dependovirus / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007763:Lac Operon / Q000235:genetics; D030781:Organisms, Genetically Modified; D019076:Transgenes / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001616:beta-Galactosidase