3D RNA-scaffolded wireframe origami.

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Biology

Background:

• Hybrid RNA:DNA origami, using RNA scaffolds and DNA oligos, offers potential for mRNA delivery and RNA structure studies but remains underexplored.
• Existing methods have limitations in fabricating complex nanostructures with long RNA molecules.

Purpose of the Study:

• To investigate design principles for three-dimensional (3D) wireframe RNA-scaffolded origami.
• To demonstrate the fabrication of diverse polyhedral nanostructures using various functional RNA scaffolds.

Main Methods:

• Computational design of RNA-scaffolded origami nanostructures.
• Fabrication of polyhedra (tetrahedra, octahedra, pentagonal bipyramids) using messenger RNA, M13 transcript RNA, and 23S ribosomal RNA.
• Characterization of secondary and tertiary structures using dimethyl sulfate mutational profiling and cryo-electron microscopy.

Main Results:

• Successful design and fabrication of diverse 3D wireframe origami polyhedra using long RNA scaffolds.
• Demonstrated ability to create complex shapes like tetrahedra, octahedra, and pentagonal bipyramids.
• Characterization provided insights into both global and local base-level structures of the RNA origami.

Conclusions:

• A top-down sequence design strategy enables the use of long RNAs as functional scaffolds for complex wireframe origami.
• This approach expands the potential of RNA origami for applications in biomedical delivery and synthetic biology.
• The study provides a framework for designing and creating sophisticated RNA-based nanostructures.