Preparation and Biological Properties of Oligonucleotide-Functionalized Virus-like Particles

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Biology

Background:

• Oligonucleotides are key for programming molecular function and assembly.
• Spherical nucleic acids (SNAs) on nanoparticle scaffolds exhibit unique properties.
• Virus-like particles (VLPs) are potential scaffolds for biomolecule presentation.

Purpose of the Study:

• To investigate if grafting oligonucleotides onto Qβ virus-like particles via click chemistry imparts SNA-like behavior.
• To assess the feasibility of using copper-binding ligands to facilitate oligonucleotide conjugation without substrate degradation.
• To evaluate the potential of these functionalized VLPs for nanoparticle-mediated delivery.

Main Methods:

• Utilized copper-catalyzed azide-alkyne cycloaddition (click chemistry) to attach oligonucleotides to Qβ VLPs.
• Optimized conjugation reactions using a small-molecule fluorogenic reporter.
• Synthesized polyvalent protein nanoparticle-oligonucleotide conjugates.

Main Results:

• The functionalized VLPs demonstrated enhanced cellular uptake, characteristic of SNAs.
• Particles showed increased protection against nuclease-mediated oligonucleotide cleavage.
• These constructs exhibited similar liver residence times and reduced antibody titers compared to unmodified particles, indicating immune shielding.

Conclusions:

• Oligonucleotide-functionalized Qβ VLPs can be synthesized using click chemistry.
• These novel constructs display SNA-like properties, including improved cellular uptake and nuclease resistance.
• Functionalized VLPs represent a viable platform for protein nanoparticle-mediated delivery of oligonucleotides and other functional molecules.