Functional patterning of DNA origami by parallel enzymatic modification.

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Biology

Background:

• DNA origami enables precise nanoscale construction.
• Functionalizing DNA origami surfaces is crucial for advanced applications.
• Existing methods for DNA origami modification can be complex or costly.

Purpose of the Study:

• To develop a rapid and cost-effective method for nanoscale patterning of functional molecules on DNA origami.
• To enable versatile surface modification of DNA origami with various functional groups.
• To expand the capabilities for creating diverse modified DNA origami structures.

Main Methods:

• Enzymatic transfer of functionalized dideoxynucleotides to DNA oligonucleotides on a DNA origami scaffold.
• Utilizing a single-tube reaction for efficient modification.
• Positioning modified oligonucleotides approximately 6 nm apart on a 70 × 100 nm(2) rectangular DNA origami.

Main Results:

• Demonstrated successful nanoscale patterning of functional molecules on DNA origami.
• Achieved high yields of oligonucleotide labeling and incorporation into the DNA origami.
• Verified modifications using gel electrophoresis, HPLC, mass spectrometry, AFM, and fluorescence measurements.
• Showcased the ability to introduce chemical handles, fluorescent dyes, and protein-binding ligands.

Conclusions:

• The developed enzymatic technique is rapid, cost-effective, and efficient for DNA origami surface functionalization.
• This method allows for the creation of diverse modified DNA origami from a single set of staple strands.
• The technique significantly expands the toolbox for designing and constructing custom DNA origami nanostructures.