Determinants of ligand-functionalized DNA nanostructure-cell interactions

Abstract

Synthesis of ligand-functionalized nanomaterials with control over size, shape and ligand orientation, facilitates the design of tailored nanomedicines for therapeutic purposes. DNA nanotechnology has emerged as a powerful tool to rationally construct two- and three-dimensional nanostructures, enabling site-specific incorporation of protein ligands with control over stoichiometry and orientation. To efficiently target cell surface receptors, exploration of the parameters that modulate cellular accessibility of these nanostructures is essential. In this study we systematically investigate tunable design parameters of antibody-functionalized DNA nanostructures binding to therapeutically relevant receptors. We show that, although the native affinity of antibody-functionalized DNA nanostructures remains unaltered, the absolute number of bound surface receptors is lower compared to soluble antibodies and is mainly governed by nanostructure size and DNA handle location. The obtained results provide key insights in the ability of ligand-functionalized DNA nanostructures to bind surface receptors and yields design rules for optimal cellular targeting.

Precision and Accuracy of Receptor Quantification on Synthetic and Biological Surfaces Using DNA-PAINT

“…Plotting (Figure 4d) the densities of 50 individual cells for each one of the cell lines, the average of each population can be seen: Low = 0.08 receptors/μm 2 , Int = 0.14 receptors/μm 2 , and High = 0.25 receptors/μm 2 . This is in accordance with flow cytometry measurements on these cells published by Cremers et al 41 From this, we can conclude that the approach and data analysis procedure are not sensitive to unspecific interactions and artifacts evidenced by the variation in the counted number of receptors between the control and the expressing cell lines (Figure 4d NC), even at a low receptor expression. This leads to high sensitivity and opens the door to detect low levels of expression due to the high specificity of the method.…”

Peptide-DNA origami as a cryoprotectant for cell preservation

“…The type, density, and arrangement of antifreezing peptides on DNA nanostructure are important design factors for its cryoprotective function as well. In addition, our system can have the potential to integrate other biological functions of DNA nanostructures such as intracellular delivery and receptor-specific binding mechanisms ( 28 , 51 ). In terms of cost-effective and mass production of cryoprotective DNA-based nanostructures, bacteriophage-based production of DNA can be adopted to markedly reduce the fabrication cost ( 52 ).…”

Optically Responsive Protein Coating of DNA Origami for Triggered Antigen Targeting

“…22 These targeting agents can be site-specifically attached, ensuring an optimal interaction with their receptor which was found to be dependent on the origami shape and orientation. 23 Antibody− antigen interactions can be furthermore studied by immobilization of small-molecule antigens onto the DNA origami surface 24 and can be applied for triggering a conformation change of the origami which is exploited for cargo display, such as drug molecules. 25,26 Currently, targeting is widely employed for the development of DNA-based tools for treatment of cancer.…”