# Resolving DNA origami structural integrity and pharmacokinetics in vivo

## TL;DR

A new method called PLASTIQ allows researchers to track DNA origami structures in the body with high precision, revealing how they degrade and how modifications like PEGylation can slow this process.

## Contribution

The introduction of PLASTIQ, a label-free proximity ligation assay for quantifying DNA origami structural integrity in vivo with high sensitivity and single-helix resolution.

## Key findings

- PLASTIQ can detect DNA origami structures in as little as 1 µl of blood with a detection limit of 0.01 fM.
- PEGylation significantly slows the degradation of DNA origami in a murine model.
- Internal DNA helices in a double-layered origami structure degrade more slowly than outer helices.

## Abstract

DNA origami holds great potential for advancing therapeutics, but the lack of methods for the precise assessment of structural integrity in vivo prevents its translation. Here we introduce proximity ligation assay for structural tracking and integrity quantification (PLASTIQ) for resolving origami structural integrity with only 1 µl of blood sample and with a detection limit of 0.01 fM. Through PLASTIQ, we could observe and quantify the dynamics of DNA origami degradation during blood circulation and evaluate the effectiveness of PEGylation for slowing this process in a murine model. Additionally, by using a double-layered barrel-like origami structure, we found distinct degradation kinetics of DNA helices depending on their specific location, revealing the slower degradation of internal helices compared with the outer ones. Our results suggest that PLASTIQ offers a quantitative approach for assessing DNA origami integrity in vivo by longitudinal sampling, providing dynamic pharmaceutical-level insights for accelerating the development of DNA-nanostructure-based therapeutic molecules and drugs.

A label-free, DNA-based proximity ligation assay that uses ligatable staple pairs enables the longitudinal quantification of DNA origami structural stability dynamics in vivo, with single-helix resolution for both wireframe and lattice designs.