Reprogramming the Dynamin 2 mRNA by Spliceosome-mediated RNA Trans-splicing

Area of Science:

• Molecular Biology
• Genetics
• Biochemistry

Background:

• Dynamin 2 (DNM2) is a GTPase crucial for cellular processes, and its mutations cause autosomal dominant centronuclear myopathy.
• This rare congenital myopathy leads to skeletal muscle weakness and characteristic histopathological changes.
• Currently, no curative treatments exist for DNM2-related centronuclear myopathy.

Purpose of the Study:

• To evaluate Spliceosome-Mediated RNA Trans-splicing (SMART) as a therapeutic strategy for DNM2-related centronuclear myopathy.
• To investigate the efficacy and safety of different trans-splicing strategies (3' vs. 5') for reprogramming Dnm2-mRNA.

Main Methods:

• In vitro and in vivo experiments in mice were conducted using SMART technology.
• Both classical 3'-trans-splicing and an alternative 5'-trans-splicing strategy were tested.
• Trans-splicing efficiency and potential toxicity of pre-trans-splicing molecules were assessed.

Main Results:

• The classical 3'-trans-splicing strategy exhibited toxicity and low in vivo efficiency.
• An alternative 5'-trans-splicing strategy successfully overcame toxicity and enhanced trans-splicing frequency.
• Successful detection of trans-splicing events at both mRNA and protein levels was achieved in vitro and in vivo.

Conclusions:

• SMART technology holds potential for reprogramming mutated Dnm2-mRNA in DNM2-related centronuclear myopathy.
• The 5'-trans-splicing approach appears more promising due to reduced toxicity and improved efficiency.
• Further preclinical investigation is crucial to address potential toxicity of molecular tools used in SMART therapy.