Generation and Characterization of a Transgenic Mouse Carrying a Functional Human β-Globin Gene with the IVSI-6 Thalassemia Mutation
Analysis of transgenic protein synthesis in the TG-β-IVSI-6 mouse model. (a) Western blotting was employed to determine the presence of human β-globin in the blood of wild-type (wt), hemizygous (hemi), and homozygous (homo) mice by using a human β-globin specific primary antibody (A). Human adult A0 hemoglobin was used as migration reference. (b) Native western blotting was performed by using a human β-globin specific primary antibody (A) and by employing, as a template, wild-type (wt), hemizygous (hemi), or homozygous (homo) mouse blood, and cell extracts obtained from MEL cells either infected with T9W lentiviral vector and treated with DMSO (T9W) or treated with DMSO only (−). Red Ponceau staining was used to verify that an equal amount of sample was loaded in each well and to verify the transfer to the membrane ((B) in panels (a) and (b)). (c, d) Capillary electrophoresis of whole blood from wild-type (c) and homozygous TG-β-IVSI-6 (d) mice. Peaks generated by murine hemoglobins and hybrid -globin2/-globin2 hemoglobin are indicated; a.u., arbitrary units.
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