Oligonucleotide⁻Palladacycle Conjugates as Splice-Correcting Agents

Area of Science:

• Oligonucleotide chemistry
• Medicinal chemistry
• Molecular biology

Background:

• Oligonucleotides are key in gene regulation and therapeutics.
• Chemical modifications enhance oligonucleotide stability and function.
• Cyclopalladated compounds offer unique chemical properties.

Purpose of the Study:

• To synthesize and characterize 2'-O-Methylribo phosphorothioate oligonucleotides with cyclopalladated benzylamine conjugates.
• To evaluate the hybridization properties and splice-switching activity of these modified oligonucleotides.
• To assess the in vitro toxicity and therapeutic potential of the modified oligonucleotides.

Main Methods:

• Synthesis of modified oligonucleotides with 5'-terminal cyclopalladated benzylamine groups.
• UV melting experiments to determine duplex stability.
• Splice-switching assays in human cancer cell lines (HeLa Luc/705, HuH7_705, U-2 OS_705).

Main Results:

• Modified oligonucleotides exhibited melting temperatures approximately 5 °C lower than unmodified counterparts.
• Cyclopalladated oligonucleotides demonstrated efficient splice-correction in HeLa Luc/705 cells.
• No significant toxicity was observed in the tested cell lines.

Conclusions:

• Covalently metalated oligonucleotides can be effectively used as splice-correcting agents.
• The cyclopalladated modification enhances therapeutic potential without inducing cellular toxicity.
• This study validates the feasibility of metalated oligonucleotides as novel therapeutic agents.