Building expanded structures from tetrahedral DNA branching elements, RNA and TMV protein

Area of Science:

• Nanotechnology
• Biomaterials Science
• Synthetic Biology

Background:

• Tobacco mosaic virus-like particles (TLPs) offer a versatile platform for creating novel nanomaterials.
• Self-assembly of nucleoprotein structures requires precise control over molecular interactions.

Purpose of the Study:

• To develop a novel method for constructing complex 3D nucleoprotein architectures.
• To combine organic synthesis, enzymatic ligation, and nucleic acid scaffolding for controlled self-assembly.

Main Methods:

• Chemical and enzymatic ligation were used to assemble DNA-branched elements and RNA scaffolds.
• Tetrahedral cores with phosphorylated dinucleotide arms were synthesized and coupled to DNA linkers.
• RNA scaffolds were incorporated into TLPs before or after ligation with branched elements.
• In situ growth of protein-coated tubes and ligation of pre-fabricated tubes were employed.

Main Results:

• Novel four-armed nanoobjects were successfully synthesized, demonstrating interconnected nanotube domains.
• The structures exhibit controlled assembly of TMV coat protein (CP) around RNA scaffolds.
• Two distinct length classes of nanotube domains were achieved, with 70 CP subunits per 10 nm.
• The method allows for precise control over the formation of 3D nucleoprotein architectures.

Conclusions:

• A unique combination of synthetic chemistry, enzymatic modification, and nucleic acid-guided assembly was demonstrated.
• The developed route provides access to a new class of 3D nucleoprotein architectures with polyvalent protein elements.
• This approach holds promise for creating supramolecular systems with tunable functionalities for advanced applications.