Double-stranded RNA-binding artificial cationic oligosaccharides stabilizing siRNAs with a low N/P ratio

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Delivery

Background:

• Short interfering RNAs (siRNAs) are crucial for RNA interference (RNAi) therapeutics but require stabilization.
• Effective stabilization of double-stranded RNA (dsRNA) is essential for developing robust nucleic acid-based drugs.

Purpose of the Study:

• To develop novel dsRNA-binding molecules for enhanced thermodynamic and biological stabilization of nucleic acids.
• To evaluate the binding affinity and protective capabilities of synthesized β-(1→4)-Linked-2,6-diamino-2,6-dideoxy-d-galactopyranose oligomers (ODAGals) with dsRNAs.

Main Methods:

• Synthesis of ODAGal oligomers.
• Fluorescence anisotropy measurements to determine binding affinity (Kd).
• UV melting experiments to assess thermodynamic stabilization.
• RNase A cleavage assays to evaluate protection of siRNAs.

Main Results:

• ODAGal 3mer and 4mer showed strong binding to dsRNAs with Kd < 0.02 μM.
• ODAGal binding significantly stabilized dsRNA duplexes, confirmed by UV melting.
• The ODAGal 4mer provided substantial protection to siRNAs against RNase A degradation at low N/P ratios.

Conclusions:

• ODAGals demonstrate potent binding and stabilizing effects on dsRNAs.
• ODAGals offer significant protection to siRNAs, indicating their potential as carriers.
• These findings highlight ODAGals as promising candidates for stabilizing dsRNA-based therapeutics like RNAi drugs.