case_images: PMC10694253_01
case_images: A hemizygous splicing mutation in the DMD gene identified in the boy with DMD. (A) Clinical signs of the patient. i and ii, muscle weakness and calf pseudohypertrophy. (B) Pedigree structure in the DMD family. The black square with black arrow indicates the patient. (C) Sanger sequencing verified the mutation of c.5912_5922 + 19delinsATGTATG in this family. The black and red dotted boxes respectively represent the mutation positions of the unaffected and affected members.
case_images: fped-11-1261318-g001.jpg
case_images: PMC10694253_02_fped-11-1261318-g001.jpg
case_images: F1
case_images: The results verified that the hemizygous mutation was inherited from his mother, which was passed on from his grandmother, while his father and grandfather did not carry this mutation (Figures 1B,C).
case_images: PMC10694253_02
case_images: A hemizygous splicing mutation in the DMD gene identified in the boy with DMD. (A) Clinical signs of the patient. i and ii, muscle weakness and calf pseudohypertrophy. (B) Pedigree structure in the DMD family. The black square with black arrow indicates the patient. (C) Sanger sequencing verified the mutation of c.5912_5922 + 19delinsATGTATG in this family. The black and red dotted boxes respectively represent the mutation positions of the unaffected and affected members.
case_images: fped-11-1261318-g001.jpg
case_images: PMC10694253_02_fped-11-1261318-g001.jpg
case_images: F1
case_images: The results verified that the hemizygous mutation was inherited from his mother, which was passed on from his grandmother, while his father and grandfather did not carry this mutation (Figures 1B,C).
case_images: Pathogenic analysis of the hemizygous splicing mutation in DMD by minigene assay. (A) Electrophoretogram of RT-PCR products. Lane 1, marker; 2, the empty plasmid (EP) including SD6 and SA2; 3, the mutant plasmid (MT), 4, the wild-type plasmid (WT); 5, blank control (BL). (B) Schematic of the splicing models in this DMD mutation. (C) Sequencing result of the mutated cDNA showing the aberrant splicing of intron 41. (D) Schematic view of the splicing events in this DMD mutation.
case_images: fped-11-1261318-g002.jpg
case_images: PMC10694253_02_fped-11-1261318-g002.jpg
case_images: F2
case_images: Gel electrophoresis revealed that the WT plasmid produced two fragments, indicating two splicing events (Figure 2A). The 446 bp fragment corresponded to exon 41 and the 599 bp fragment corresponded to exon 40 and exon 41, as confirmed by Sanger Sequencing (Figures 2B,C). Cells transfected with the mutant c.5912_5922 + 19delinsATGTATG plasmids also produced two fragments, each nearly equivalent to the WT (Figure 2A). However, subsequent Sanger sequencing revealed that the two mutant fragments exhibited a deletion of 11 bp (TTGCACAAATT) in exon 41 and a retention of 12 bp (ATGTATGCCCAC) in intron 41 (Figures 2B,C). Taken together, these findings suggest that the mutation in the DMD gene produced an aberrantly spliced cDNA (Figure 2D), which accounts for DMD presentation.