mRNA-Based Solution for 47, 48, 51, and 52 Dystrophin Exon Deletions: DMD Patient-Donate Primer Cells (In Vitro) and Transgenic Mice Experimental Study (In Vivo)

Background/Objectives: Duchenne muscular dystrophy (DMD) is a genetic disorder caused by mutations in the dystrophin gene. DMD is characterized by exon deletions in about 76% of cases, with common deletions in exons 47, 48, 51, and 52. We evaluated the effectiveness of an mRNA-based therapy targeting these exon deletions, which are frequently seen in DMD patients. Methods: The current study involved two protocols: 1. applying the therapy to cells from patients diagnosed with DMD, and 2. applying the therapy to genetically modified transgenic mdx/d2 mice. After treatment, dystro-phin was detected in all experimental groups. Results: Our study showed that, both in vitro and in vivo analyses demonstrated that the mRNA-based therapy successfully restored dystrophin expression in dystrophic muscle cells and tissues. Gene expression analysis, together with protein-level assessments, including Western blot, immunoflu-orescence (IF), ELISA, and immunohistochemistry (IHC), confirmed a significant in-crease in dystrophin levels in the treated groups compared with the control group. In addition to dystrophin restoration, other key sarcolemmal proteins involved in main-taining muscle membrane stability, such as γ-sarcoglycan, β-dystroglycan, and β-actin, were also highly expressed. These findings suggest an overall improvement in muscle cell membrane integrity. Consistent with the molecular results, behavioral analyses performed in the animal model revealed significant functional improvements, includ-ing enhanced mobility, motor coordination, longer walking and resting durations, and a reduced risk of falls. Overall, the results indicate that mRNA-mediated dystrophin replacement improves both muscle structural integrity and functional performance. Conclusions: Our study proved that the mRNA complex successfully produced func-tional dystrophin in transgenic mdx/d2 mice without causing allergic reactions or damage to the kidney, intestines, muscles, or brain.