Abstract
DNA origami nanocarriers have emerged as a promising tool for many biomedical applications, such as biosensing, targeted drug delivery, and cancer immunotherapy. These highly programmable nanoarchitectures are assembled into any shape or size with nanoscale precision by folding a single-stranded DNA scaffold with short complementary oligonucleotides. The standard scaffold strand used to fold DNA origami nanocarriers is usually the M13mp18 bacteriophage’s circular single-stranded DNA genome with limited design flexibility in terms of the sequence and size of the final objects. However, with the recent progress in automated DNA origami design—allowing for increasing structural complexity—and the growing number of applications, the need for scalable methods to produce custom scaffolds has become crucial to overcome the limitations of traditional methods for scaffold production. Improved scaffold synthesis strategies will help to broaden the use of DNA origami for more biomedical applications. To this end, several techniques have been developed in recent years for the scalable synthesis of single stranded DNA scaffolds with custom lengths and sequences. This review focuses on these methods and the progress that has been made to address the challenges confronting custom scaffold production for large-scale DNA origami assembly.
Highlights
In all known living organisms, DNA molecules are responsible for storing and carrying genetic information [1]
The potential of DNA origami nanoparticles has been demonstrated through many successful biomedical applications, including drug delivery, vaccine platform development, and cancer therapy [19,20,21,22,25,26]
We have presented the various strategies that have been developed, or are under development, to synthesize long strands of DNA (ssDNA) scaffolds for DNA origami folding
Summary
In all known living organisms, DNA molecules are responsible for storing and carrying genetic information [1]. DNA origami nanoparticles aretheassembled by folding a longdiscrete ssDNA scaffold strand with an excess method rapidly became strategy of choice for synthesizing nanometer-scale particles, notably enabling the assembly of custom complex. Enabling the automated design of complex nanostructures with any shape7249 or size—and the increasing the M13mp bacteriophage’s genome—a commercially available nucleotide (nt)-long circular number of biomedical applications [43,44,45] have led to the increased complexity and size of designed single strand of DNA—which can readily be used to assemble nanoparticles in a 10 to 100 nm DNA origami. This review focuses on existing and emerging techniques for the synthesis of ssDNA scaffolds for DNA origami folding It describes the various bacteriophage production methods, enzymatic synthesis strategies, and highlights promising new approaches to further develop the existing toolbox for scaffold synthesis. The methodologies, yields, functionality, and limitations of each method are presented
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