DNA nanotechnology-based biomolecular assembly and imaging
Arranging biomacromolecules in a specific order and orientation on the nanoscale has long been a prominent research topic.Significant progress has been made in understanding intermolecular interactions within bionetworks and elucidating biosystem mechanisms.DNA nanostructures,serving as ideal scaffolds,have been extensively employed to achieve precise arrangement of diverse biomolecules at the nanoscale due to their well-defined geometry,programmability,addressability at the nanometer scale,functionalization capabilities and excellent biocompatibility.The advent of DNA origami has further enhanced the ability to design intricate and accurate DNA frameworks.This technique utilizes numerous short DNA strands,called staples,to fold a longer DNA strand,known as the scaffold,enabling the creation of more structurally complex and precise DNA architectures.This review is mainly structured into four main sections.Firstly,we provide a comprehensive overview of the various methods employed for the three key steps involved in assembling DNA nanostructures with biomolecules.These steps encompass the conjugation of oligonucleotides and biomolecules,the hybridization of DNA nanostructures with biomolecules,and the purification of DNA nanostructures-biomolecule complexes.Secondly,we briefly summarize the relevant applications of DNA nanostructures-biomolecules self-assembly,with a primary focus on the construction of enzyme cascade reaction systems.The combination of enzyme cascades and addressable DNA assembly has proven highly effective in creating stable artificial multi-enzyme complexes.This integration allows for precise control over the position,shape,and composition of the complexes,enabling efficient manipulation of enzyme distance,substrate channeling,and compartmentalization.Thirdly,we focus on the application of DNA nanostructures in diverse imaging techniques,including atomic force microscopy(AFM),fluorescence imaging,and cryo-electron microscopy(cryo-EM).DNA nanostructure offers distinct advantages in advancing bioimaging modalities,such as enabling multiplexed imaging,facilitating logical responsive imaging,and enabling protein structural analysis.While AFM has been extensively employed for the characterization of various biomolecules,it still has limitations in direct observation and comparison of freely dispersed molecules.However,by utilizing DNA nanostructures as a platform for loading molecules,it becomes possible to realize the visualization of dynamic intermolecular interactions using AFM.Besides,through precisely manipulating the number and combination of selected fluorophores,DNA nanostructures provide a means to mitigate the issue of spectral overlap among fluorophore colors.This precise control allows for tuning the overall color of probes,effectively overcoming spectrum overlapping and enabling the generation of distinct and distinguishable colors.Moreover,DNA nanostructures show potential in assisting the determination of protein orientation during cryo-electron microscopy(cryo-EM)imaging and potentially mitigating issues such as protein unfolding or nonrandom absorption at the water-air interface.The final section of this review discusses the technical challenges that need to be addressed to enhance the utilization of DNA nanoscaffolds,for example achieving precise orientation control of biomolecules on DNA scaffolds,optimizing the stability of the overall complexes to achieve in vivo functionality and overcoming technological bottlenecks that impede the application of DNA nanoscaffolds in cryo-EM imaging.
DNA nanotechnologyDNA origamibiomolecule assemblybiomolecule imaging
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维普
