Tunable Nanoscale Cages from Self-Assembling DNA and Protein Building Blocks.

Area of Science:

• Nanotechnology
• Molecular Engineering
• Biomaterials Science

Background:

• Proteins and DNA are key building blocks for nanotechnology, enabling tunable nanoscale cage construction.
• Existing protein- and DNA-based cages have inherent limitations.
• Hybrid approaches are needed to leverage the advantages of both molecular systems.

Purpose of the Study:

• To develop a novel method for constructing three-dimensional (3D) nanoscale cages using both protein and DNA components.
• To create tunable hybrid cages with controllable dimensions.
• To demonstrate the versatility and generality of the protein-DNA conjugation strategy.

Main Methods:

• Covalent conjugation of a homotrimeric protein (KDPG aldolase) with single-stranded DNA handles via site-directed mutagenesis and functionalization of cysteine residues.
• Co-assembly of protein-DNA building blocks with complementary triangular DNA structures to form tetrahedral cages.
• Characterization of hybrid cage structures using purification and analytical techniques.
• Demonstration of cage modification using click chemistry and noncanonical amino acids.

Main Results:

• Successfully constructed hybrid protein-DNA tetrahedral cages with tunable dimensions (∼10 nm and ∼14 nm).
• Confirmed the 3D structure and integrity of the assembled hybrid cages.
• Demonstrated the adaptability of the method for further functionalization using click chemistry and noncanonical amino acids.

Conclusions:

• A novel and versatile method for creating hybrid protein-DNA nanoscale cages has been developed.
• This approach combines the advantages of protein and DNA nanotechnology, overcoming individual limitations.
• The resulting tunable hybrid cages hold significant potential for applications in targeted delivery, structural biology, biomedicine, and catalysis.