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Purification of DNA Nanoparticles Using Photocleavable Biotin Tethers.

Heather R Everson¹, Kayla Neyra¹, Dylan V Scarton^2,3

¹Department of Chemistry, Case Western Reserve University, Cleveland, Ohio 44106, United States.

ACS Applied Materials & Interfaces

|April 18, 2024

Summary

Researchers developed a simple, single-step purification method for DNA origami nanoparticles (DNA NPs) using photocleavable biotin tethers and UV light. This technique efficiently isolates custom DNA NPs, improving yield and purity for various applications.

Keywords:

DNA nanotechnology DNA origami asymmetric polymerase chain reaction biotin magnetic beads nanostructures purification

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Area of Science:

Nanotechnology
Biochemistry
Materials Science

Background:

Self-assembled deoxyribonucleic acid (DNA) origami nanoparticles (DNA NPs) are increasingly utilized, with single-stranded DNA (ssDNA) scaffolds derived from viral genomes or enzymatic production gaining prominence.
Current purification of DNA NPs from assembly mixes is a multistep process, involving separation of the ssDNA scaffold and then isolation of the assembled DNA NPs, delaying downstream applications.

Purpose of the Study:

To develop and demonstrate a streamlined, single-step purification method for DNA origami nanoparticles (DNA NPs) utilizing custom or M13-derived scaffolds.
To enhance the efficiency and accessibility of DNA NP purification for broader research and application.

Main Methods:

A novel purification strategy employing photocleavable biotin tethers was implemented for DNA NPs.
Purification was achieved in a single step using an inexpensive ultraviolet (UV) lamp to cleave the tethers.

Main Results:

The single-step purification method yielded DNA NPs with up to 90% efficiency and high purity.
The technique proved versatile, successfully separating different DNA NP architectures, including multihelix bundles and wireframe polyhedra.

Conclusions:

Photocleavable biotin tethers offer an efficient and cost-effective single-step purification solution for DNA origami nanoparticles.
This simplified purification process accelerates the readiness of DNA NPs for characterization and diverse downstream applications.