Resolving DNA origami structural integrity and pharmacokinetics in vivo

Area of Science:

• Biotechnology
• Nanomedicine
• Molecular Engineering

Background:

• DNA origami shows therapeutic promise but lacks in vivo structural integrity assessment methods.
• This limitation hinders the clinical translation of DNA nanostructures for drug delivery.

Purpose of the Study:

• To introduce a novel method for quantifying DNA origami structural integrity in vivo.
• To enable dynamic monitoring of DNA origami degradation and evaluate stabilization strategies.

Main Methods:

• Proximity ligation assay for structural tracking and integrity quantification (PLASTIQ) was developed.
• PLASTIQ utilizes a small blood volume (1 µl) with high sensitivity (0.01 fM detection limit).
• The method was applied in a murine model to track DNA origami degradation and assess PEGylation efficacy.

Main Results:

• PLASTIQ successfully quantified DNA origami degradation dynamics in circulating blood.
• PEGylation was observed to significantly slow down DNA origami degradation in vivo.
• Distinct degradation rates were found for internal versus external DNA helices in a barrel-like structure.

Conclusions:

• PLASTIQ provides a quantitative, longitudinal approach for assessing in vivo DNA origami integrity.
• This method offers pharmaceutical-level insights crucial for accelerating DNA nanostructure-based therapeutic development.
• Understanding degradation kinetics informs the design of more stable and effective DNA nanomedicines.