Small Molecule‐Templated DNA Hydrogel with Record Stiffness Integrates and Releases DNA Nanostructures and Gene Silencing Nucleic Acids

Deoxyribonucleic acid (DNA) hydrogels are a unique class of programmable, biocompatible materials able to respond to complex stimuli, making them valuable in drug delivery, analyte detection, cell growth, and shape‐memory materials. However, unmodified DNA hydrogels in the literature are very soft, rarely reaching a storage modulus of 10(3) Pa, and they lack functionality, limiting their applications. Here, a DNA/small‐molecule motif to create stiff hydrogels from unmodified DNA, reaching 10(5) Pa in storage modulus is used. The motif consists of an interaction between polyadenine and cyanuric acid—which has 3‐thymine like faces—into multimicrometer supramolecular fibers. The mechanical properties of these hydrogels are readily tuned, they are self‐healing and thixotropic. They integrate a high density of small, nontoxic molecules, and are functionalized simply by varying the molecule sidechain. They respond to three independent stimuli, including a small molecule stimulus. These stimuli are used to integrate and release DNA wireframe and DNA origami nanostructures within the hydrogel. The hydrogel is applied as an injectable delivery vector, releasing an antisense oligonucleotide in cells, and increasing its gene silencing efficacy. This work provides tunable, stimuli‐responsive, exceptionally stiff all‐DNA hydrogels from simple sequences, extending these materials’ capabilities.