Regulated high level expression of a human gamma-globin gene introduced into erythroid cells by an adeno-associated virus vector.
AUTOR(ES)
Walsh, C E
RESUMO
Gene therapy of severe hemoglobinopathies will require high-level expression of a transferred globin gene in erythroid cells. Distant regulatory elements flanking the beta-globin gene cluster, the locus control region, are needed for appropriate expression. We have explored the use of a human parvovirus, the adeno-associated virus (AAV), for globin gene transfer. The human A gamma-globin gene, linked to hypersensitivity site 2 from the locus control region of the beta-globin gene cluster, was subcloned into a plasmid (psub201) containing the AAV inverted terminal repeats. This construct was cotransfected with a helper plasmid containing trans-acting AAV genes into human 293 cells that had been infected with adenovirus. The recombinant AAV vector containing hypersensitivity site 2 stably introduced on average one or two unrearranged proviral copies into human K562 erythroleukemia cells. The transferred globin gene exhibited normal regulation upon hemin induction of erythroid maturation and was expressed at a level equivalent to a native chromosomal A gamma-globin gene.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=49685Documentos Relacionados
- Recombinant adeno-associated virus (rAAV)-mediated expression of a human gamma-globin gene in human progenitor-derived erythroid cells.
- Efficient long-term gene transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector.
- Phenotypic correction of Fanconi anemia in human hematopoietic cells with a recombinant adeno-associated virus vector.
- Role for highly regulated rep gene expression in adeno-associated virus vector production.
- Nuclear protein factors and erythroid transcription of the human A gamma-globin gene.