title: Long-term expression of the human beta-globin gene after retroviral transfer into pluripotent hematopoietic stem cells of the mouse.
abstract: We have studied the regulation of the human beta-globin gene after retroviral transfer into a variety of transformed and normal hematopoietic cells. After transfer into murine erythroleukemia cells (MEL) expression from the human beta-globin gene responds to inducers of erythroid maturation in parallel to the endogenous murine globin genes. After infection of human BFU-E, RNA expression from the virally-transferred beta-globin gene was measured at 2.5%-5% of the endogenous beta-globin level. The most improved globin vectors can transfer the human beta-globin gene into pluripotent hematopoietic stem cells in mouse bone marrow. Mice reconstituted with infected marrow show human beta-globin RNA and protein expression in peripheral blood cells for over 4 months. In these animals, both myeloid and lymphoid cells carry the integrated provirus at a level of about 1 copy per cell. In serial transplantation experiments, bone marrow from these animals is capable of repopulating secondary and tertiary recipient animals which go on to show long-term human beta-globin expression. Retroviral vectors thus provide a practical way to refine models of globin gene regulation through in vivo tests and to evaluate the feasibility of protocols for gene addition therapy.
authors.affiliation: Department of Molecular Medicine, Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
authors.forename: R E
authors.initials: RE
authors.lastname: Gelinas
authors.forename: M A
authors.initials: MA
authors.lastname: Bender
authors.forename: A D
authors.initials: AD
authors.lastname: Miller
authors.forename: U
authors.initials: U
authors.lastname: Novak
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation; D002478:Cells, Cultured; D002678:Chimera; D015672:Erythroid Precursor Cells / Q000378:metabolism; D005786:Gene Expression Regulation; D005796:Genes; D005822:Genetic Vectors; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006085:Graft Survival; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D007438:Introns; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D051379:Mice; D014411:Neoplastic Stem Cells / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012190:Retroviridae; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D005914:Globins