Construction of T‐Motif‐Based DNA Nanostructures through Enzymatic Reactions

Abstract

The most common way to fabricate DNA nanostructures is to mix individually synthesized DNA oligomers in one pot. However, if DNA nanostructures could be produced through enzymatic reactions, they could be applied in various environments, including in vivo. Herein, an enzymatic method developed to construct a DNA nanostructure from a simple motif called a T-motif is reported. A long, repeated structure was replicated from a circular template by rolling circle amplification and then cleaved into T-motif segments by restriction enzymes. These motifs have been successfully assembled into a ladder-like nanostructure without purification or controlled annealing. This approach is widely applicable to constructing a variety of DNA nanostructures through enzymatic reactions.

DNA nanotechnology provides an avenue for the construction of programmable dynamic molecular systems

“…It should be noted that various techniques, such as induction of changes in ionic strength [7] and/or pH [8], light-irradiation [66,67], and addition of fuel molecules [68,69], can be employed to trigger self-assembly of DNA nanostructures, enabling the construction of various artificial signal-responsive molecular systems that exhibit deformation [67], locomotion [70], swarm behaviors [71], and production of components of supramolecular structures [72]. The recent development of reversible mechanical motion of DNA nanostructures [73] and DNA-based circuits [74] with modularity would empower DNA nanotechnology to enable designing a molecular system that possesses adaptability to environmental changes [75,76].…”

Reconfigurable T‐junction DNA Origami

“…[20] The 5' T-junction has been used to assemble 2D arrays, [20,21] polyhedra, [21,22] and RNA nanostructures. [23] It is well suited to systems designed for enzymatic [24] or enzymefree [25,26] copying. The weaker 3' T-junction has not been used as a construction motif although recent results suggest that both 5' and 3' motifs can be used.…”

Golden Gate Assembly of DNA Nanostructures

“…DNA processing enzymes have been applied to synthetic DNA constructs since the outset of DNA nanotechnology. , Because the DNA duplex segments constitute the major body of a DNA nanostructure, it is natural to use enzymes from the molecular cloning toolbox for the specific modifications of synthetic DNA constructs. − For example, polymerases can be used to fill gaps of double-stranded DNA, − restriction enzymes can be applied to cleave specific sequence motifs to create nicks for short strands in DNA nanostructures, − and ligase can also be used to seal nicks for multiple short strands to be combined covalently. ,,,− Upon enzymatic processing, new features are brought to the resulting constructs. More importantly, DNA nanostructure dynamics of a higher level of controllability and reliability induced by enzymatic treatments could lead to the precise execution of advanced computation.…”