Designer DX-tile DNAns hydrogels

Pure deoxyribonucleic acid (DNA) hydrogels synthesized via the hybridization of multi-arm DNA tiles are uniquely programmable and functionalizable biomaterials suitable for applications ranging from biosensing to cell-free protein production and soft tissue engineering. However, the full potential of the design flexibility and functionalization offered by DNA molecules has not yet been leveraged for pure DNA hydrogels, thereby limiting their range of mechanical properties and reducing their versatility and broader use. In this study, we introduce multi-arm double-crossover (DX)-tile motifs, often used in DNA nanoparticle design, to enable greater control over the hydrogel’s mechanical properties and facilitate functionalization. Specifically, we demonstrate that modifying structural design parameters, such as the arm geometry, length, valency, and linker design, allows fine control of the elastic modulus and viscoelastic properties of the hydrogels. We also show that functionalization can be performed without compromising the hydrogels’ physical properties and exhibit enhanced mechanical strength and tunable properties, compared to simple duplex-based DNA hydrogels. Furthermore, these DNA hydrogels demonstrated printability and scalability, which pave the way towards the development of novel formulations and bioinks for the rational design of soft tissue engineering scaffolds and broaden the use of DNA hydrogels for other biomedical applications.