Sprecher
Beschreibung
Self-assembled deoxyribonucleic acid (DNA) origami nanostructures (DONs) are highly versatile, possessing unique structural and mechanical properties that make them promising candidates for diverse applications such as biosensing, drug delivery, and advanced nanodevices. However, realizing their full potential often requires precise pre- or post-assembly processing/ modification for functionalization or dynamic applications. Nucleases offer a powerful toolkit for targeted and localized structural modification of DONs such as controlled cutting and joining; however, achieving this through traditional enzymatic processing remains challenging because of their topological complexity. Our research investigates the controlled action of specific and non-specific nucleases on both 2D and 3D DONs. Triangular 2D DONs (with trapezoidal domains bridged through single stranded linkers) were assembled and processed with various exonucleases in a time-lapse manner and then characterized using agarose gel electrophoresis (AGE) and atomic force microscopy (AFM). Our initial findings reveal distinct degradation patterns: non-specific nucleases (e.g., DNase I, Exonuclease III) lead to rapid, uncontrolled degradation of DONs, while Mung Bean Nuclease (MBN) demonstrated relatively selective and controlled cutting at exposed single stranded linkers and overhangs. These results highlight the potential of MBN and other specific exonucleases as powerful tools for controlled cleavage and modification of complex DNA origami nanostructures in a programmable manner. On-going work aims to further elucidate the dynamic interactions between various specific and non-specific nucleases (DNase I, Exonuclease III, MBN) and both 2D and 3D DONs for their utility in advanced DNA nanotechnology applications.
Keywords: Self-assembly; DNA origami; Enzyme reactions; Nucleases; Exonucleases; Electrophoresis; AFM.
