title: Hypoxia-induced in vivo sickling of transgenic mouse red cells.
abstract: To develop an animal model for sickle cell anemia, we have created transgenic mice that express a severe naturally occurring human sickling hemoglobin, Hb S Antilles. Due to its low solubility and oxygen affinity, Hb S Antilles has a greater propensity to cause red cell sickling than Hb S. To make transgenic animals that express a high level of Hb S Antilles, the erythroid-specific DNAse I hypersensitive site II from the human beta-globin cluster was linked independently to the human alpha 2-globin gene and to the beta S Antilles gene. Embryos were injected with both constructs simultaneously and seven transgenic mice were obtained, three of which contained both the human alpha and the human beta S Antilles transgene. After crossing the human transgenes into the mouse beta-thalassemic background a transgenic mouse line was derived in which approximately half the beta-globin chains in the murine red cells were human beta S Antilles. Deoxygenation of the transgenic red cells in vitro resulted in extensive sickling. An increase of in vivo sickling was achieved by placing these transgenic mice in a low oxygen environment. This murine model for red cell sickling should help to advance our understanding of sickle cell disease and may provide a model to test therapeutic interventions.
authors.affiliation: Division of Cell and Molecular Biology, Lawrence Berkeley Laboratory, University of California, Berkeley 94720.
authors.forename: E M
authors.initials: EM
authors.lastname: Rubin
authors.forename: H E
authors.initials: HE
authors.lastname: Witkowska
authors.forename: E
authors.initials: E
authors.lastname: Spangler
authors.forename: P
authors.initials: P
authors.lastname: Curtin
authors.forename: B H
authors.initials: BH
authors.lastname: Lubin
authors.forename: N
authors.initials: N
authors.lastname: Mohandas
authors.forename: S M
authors.initials: SM
authors.lastname: Clift
mesh_terms: D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D004195:Disease Models, Animal; D004912:Erythrocytes / Q000648:ultrastructure*; D005914:Globins / Q000235:genetics / Q000302:isolation & purification; D006451:Hemoglobin, Sickle / Q000378:metabolism*; D000860:Hypoxia / Q000503:physiopathology*; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data
chemical_list: D006451:Hemoglobin, Sickle; D005914:Globins; D004247:DNA