Development of the 12-Base Short Dimeric Myogenetic Oligodeoxynucleotide That Induces Myogenic Differentiation

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Development

Background:

• Myogenetic oligodeoxynucleotides (myoDNs) like iSN04 show promise for treating muscle wasting by inducing myogenic differentiation.
• Current myoDNs face challenges in drug efficacy and synthesis cost, necessitating sequence optimization.

Purpose of the Study:

• To develop a shorter, cost-effective myoDN with preserved myogenic activity.
• To investigate the structural basis for the function of truncated myoDNs.

Main Methods:

• Synthesis and characterization of a novel 12-base myoDN (iMyo01).
• Assay of myogenic activity in primary human myoblasts.
• Nucleolin binding studies.
• Nuclear magnetic resonance (NMR) and native polyacrylamide gel electrophoresis (PAGE).
• Computational molecular dynamics simulations.

Main Results:

• The 12-base iMyo01 demonstrated comparable myogenic activity to the 18-base iSN04.
• Both iMyo01 and iSN04 interacted with nucleolin and upregulated myogenic gene expression.
• iMyo01 forms a stable G-quadruplex structure, potentially as a homodimer, despite its short, non-telomeric sequence.
• Unlike iSN04, iMyo01 did not bind to berberine.

Conclusions:

• Aptamer truncation technology can yield shorter, bioactive nucleic acid drugs.
• iMyo01 represents a promising, optimized myoDN for potential therapeutic applications in muscle wasting diseases.
• The G-quadruplex structure formed by iMyo01 is key to its preserved bioactivity.