Generation and Characterization of a Transgenic Mouse Carrying a Functional Human β-Globin Gene with the IVSI-6 Thalassemia Mutation

Figure 9

Normal and aberrant splicing in IVSI-6 β-globin gene. (a) Schematic representation of the normal (A) and altered splicing (B, C, D) in IVSI-6 thalassemia. Grey arrows indicate the primers used to demonstrate the presence of the altered splicing. The positions of the cryptic splicing sites generated by the mutation and the respective lengths (in bp) of products obtained after PCR amplification of altered transcripts are indicated with different colours. A red star locates the IVSI-6 mutation. (b, c) Identification of aberrantly spliced transcripts in IVSI-6 patients and in the TG-β-IVSI-6 mouse model. (b) Electropherograms generated by denaturing polyacrylamide gel electrophoresis of fluorescent RT-PCR products obtained from healthy donor blood (left panel) and IVSI-6 homozygous patient blood (right panel). (c) Electropherograms obtained from a wild-type and two TG-β-IVSI-6 mice (A, B). Primers employed were HuBetaF1[6FAM]-HuBetaR (Table 2). Blue peaks indicate both alternatively spliced and canonic human transcripts.

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