DNA Origami

DNA Origami
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DNA ORIGAMI Discover the impact and multidisciplinary applications of this subfield of DNA nanotechnology DNA origami refers to the technique of assembling single-stranded DNA template molecules into target two- and three-dimensional shapes at the nanoscale. This is accomplished by annealing templates with hundreds of DNA strands and then binding them through the specific base-pairing of complementary bases. The inherent properties of these DNA molecules—molecular recognition, self-assembly, programmability, and structural predictability—has given rise to intriguing applications from drug delivery systems to uses in circuitry in plasmonic devices. The first book to examine this important subfield, DNA Origami brings together leading experts from all fields to explain the current state and future directions of this cutting-edge avenue of study. The book begins by providing a detailed examination of structural design and assembly systems and their applications. As DNA origami technology is growing in popularity in the disciplines of chemistry, materials science, physics, biophysics, biology, and medicine, interdisciplinary studies are classified and discussed in detail. In particular, the book focuses on DNA origami used for creating new functional materials (combining chemistry and materials science; DNA origami for single-molecule analysis and measurements (as applied in physics and biophysics); and DNA origami for biological detection, diagnosis and therapeutics (medical and biological applications). DNA Origami readers will also find: A complete guide for newcomers that brings together fundamental and developmental aspects of DNA origami technology Contributions by a leading team of experts that bring expert views from different angles of the structural developments and applications of DNA origami An emerging and impactful research topic that will be of interest in numerous multidisciplinary areas A helpful list of references provided at the end of each chapter to give avenues for further study Given the wide scope found in this groundbreaking work, DNA Origami is a perfect resource for nanotechnologists, biologists, biophysicists, chemists, materials scientists, medical scientists, and pharmaceutical researchers.

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Группа авторов. DNA Origami

Table of Contents

List of Tables

List of Illustrations

Guide

Pages

DNA Origami. Structures, Technology, and Applications

List of Contributors

Preface

1 DNA Origami Technology: Achievements in the Initial 10 Years

1.1 Introduction

1.1.1 DNA Nanotechnology Before the Emergence of DNA Origami

1.2 Two‐Dimensional DNA Origami

1.3 Programmed Arrangement of Multiple DNA Origami Components

1.4 Three‐Dimensional DNA Origami Structures

1.5 Modification and Functionalization of 2D DNA Origami Structures. 1.5.1 Selective Placement of Functional Nanomaterials

1.5.2 Selective Placement of Functional Molecules and Proteins via Ligands

1.5.3 Distance‐Controlled Enzyme Reactions and Photoreactions

1.6 Single‐Molecule Detection and Sensing using DNA Origami Structures. 1.6.1 Single‐Molecule RNA Detection

1.6.2 Single‐Molecule Detection of Chemical Reactions

1.6.3 Single‐Molecule Detection using Mechanical DNA Origami

1.6.4 Single‐Molecule Sensing using Mechanical DNA Origami

1.7 Application to Single Biomolecule AFM Imaging. 1.7.1 High‐Speed AFM‐Based Observation of Biomolecules

1.7.2 Visualization of DNA Structural Changes in the DNA Nanospace

1.7.3 Visualization of the Reaction Events of Enzymes and Proteins in the DNA Nanospace

1.8 Single‐Molecule Fluorescence Studies

1.8.1 Nanoscopic Ruler for Single‐Molecule Imaging

1.8.2 Kinetics of Binding and Unbinding Events and DNA‐PAINT

1.8.3 DNA Barcode Imaged by DNA‐PAINT

1.9 DNA Molecular Machines

1.9.1 DNA Assembly Line Constructed on the DNA Origami

1.9.2 DNA Spider System Constructed on the DNA Origami

1.9.3 DNA Motor System Constructed on the DNA Origami

1.10 Selective Incorporation of Nanomaterials and the Applications

1.10.1 DNA Origami Plasmonic Structure with Chirality

1.10.2 Surface‐Enhanced Fluorescence by Gold Nanoparticles and DNA Origami Structure

1.10.3 Placement of DNA Origami onto a Fabricated Solid Surface

1.11 Dynamic DNA Origami Structures Responsive to External Stimuli. 1.11.1 DNA Origami Structures Responsive to External Stimuli

1.11.2 Stimuli‐Responsive DNA Origami Plasmonic Structures

1.11.3 Photo‐Controlled DNA Origami Plasmonic Structures

1.12 Conjugation of DNA Origami to Lipid. 1.12.1 DNA Origami Channel with Gating

1.12.2 DNA Origami Templated Synthesis of Liposomes

1.13 DNA Origami for Biological Applications

1.13.1 Introduction of DNA Origami into Cells and Functional Expression

1.13.2 Drug Release Using the Properties Characteristic for DNA Origami

1.13.3 DNA Origami Structures Coated with Lipids and Polymers

1.13.4 Nanorobot with Dynamic Mechanism

1.13.5 Nanorobot Targeting Tumor In Vivo

1.14 Conclusions

References

2 Wireframe DNA Origami and Its Application as Tools for Molecular Force Generation

2.1 Introduction

2.2 Pre‐Origami Wireframe DNA Nanostructures

2.3 Hierarchical DNA Origami Wireframe

2.4 Entire DNA Origami Design

2.5 DNA Origami Wireframe as Tools for Molecular Force Application. 2.5.1 Introduction

2.5.2 Results and Discussion

2.6 Conclusions

2.6.1 Materials and Methods

References

3 Capturing Structural Switching and Self‐Assembly Events Using High‐Speed Atomic Force Microscopy

3.1 Introduction

3.2 DNA Origami Nanomachines

3.3 Ion‐Responsive Mechanical DNA Origami Devices

3.4 Photoresponsive Devices

3.5 Two‐Dimensional Self‐Assembly Processes

3.6 Sequential Self‐Assembly

3.7 Photostimulated Assembly and Disassembly

3.8 Conclusions and Perspectives

References

4 Advancement of Computer‐Aided Design Software and Simulation Tools for Nucleic Acid Nanostructures and DNA Origami

4.1 Introduction

4.2 General‐Purpose Software

4.3 Software for Designing Small DNA Nanostructures

4.4 Software for Designing DNA Origami

4.5 Software for Designing RNA Nanostructures

4.6 Software for Designing Base Sequence

4.7 Software for Simulating Nucleic Acid Nanostructures

4.8 Summary and Future Perspective

References

5 Dynamic and Mechanical Applications of DNA Nanostructures in Biophysics

5.1 Introduction

5.1.1 What Makes DNA a Good Material for Dynamic Applications

5.1.2 Rupture Forces

5.2 Applications

5.2.1 Force Spectroscopy

5.2.1.1 Utilizing the Stiffness of DNA for Force Spectroscopy

5.2.1.2 Applications that Utilize Rupture Forces

5.2.2 DNA Devices that Probe and Control DNA–DNA Interactions. 5.2.2.1 Detection

5.2.2.2 Modulation

5.2.3 DNA Devices that Respond to Biomolecules

5.2.4 DNA Devices to Study Biological Molecular Motors

5.2.5 DNA Walkers

5.2.6 DNA Computing

5.3 Tools for Quantifying DNA Devices and their Functions

5.4 Modeling and Analysis

5.5 Conclusion

References

6 Plasmonic Nanostructures Assembled by DNA Origami

6.1 Introduction

6.2 Optical Properties of the DNA Origami‐Based Plasmonic Nanostructures

6.3 Nanoparticle Functionalization with DNA

6.4 DNA Origami‐Based Plasmonic Assemblies

6.5 Surface‐Enhanced Raman Scattering (SERS) and Other Plasmonic Effects

6.6 Conclusion

Acknowledgments

References

7 Assembly of Nanoparticle Superlattices Using DNA Origamias a Template

7.1 Introduction

7.2 Gold Nanoparticles

7.2.1 Oligonucleotide‐Modified AuNPs

7.2.2 Cationic AuNPs

7.3 Formation of DNA Origami‐Assisted Superlattices

7.3.1 Superlattices Formed by Oligonucleotide‐Functionalized AuNPs

7.3.2 Superlattice Formed by Cationic AuNPs

7.4 Characterization of Assemblies

7.4.1 Electron Microscopy

7.4.2 Small‐Angle X‐ray Scattering

7.5 Conclusions and Future Perspectives

Acknowledgments

References

8 Mechanics of DNA Origami Nanoassemblies

8.1 Introduction

8.2 Analytical Tools to Investigate Mechanical Properties of Nanoassemblies

8.2.1 Optical Tweezers

8.2.2 Magnetic Tweezers

8.2.3 Atomic Force Microscopy (AFM)

8.3 Mechanical Strength of DNA Origami Structures

8.4 Applications of Origami Nanostructures by Exploiting their Mechanical Strength

8.5 Mechanochemical Properties of DNA Origami

8.6 Conclusions

References

9 3D DNA Origami as Single‐Molecule Biophysical Tools for Dissecting Molecular Motor Functions

9.1 Introduction

9.2 DNA Origami Nanospring. 9.2.1 Design of DNA Origami Nanospring

9.2.2 Nanospring Mechanical Properties

9.2.3 Application to a Myosin VI Processive Motor

9.3 DNA Origami Thick Filament Mimicking Muscle Structure. 9.3.1 Mystery of Muscle Contraction

9.3.2 Design of a DNA Origami‐Based Thick Filament

9.3.3 High‐speed AFM Observation of Force Generation by Myosin

9.3.4 High‐Speed Darkfield Imaging of Force Generation by Myosin

9.4 Perspective

References

10 Switchable DNA Origami Nanostructures and Their Applications

10.1 Introduction

10.2 Switchable Machines Constructed from DNA Origami Scaffolds

10.2.1 Chemical Triggers for Origami Scaffolds. 10.2.1.1 Triggering Origami Devices with Strand Displacement Reactions

10.2.1.2 Triggering Origami with Ion Concentration

10.2.1.3 Triggering Origami with Molecular Species

10.2.2 Physical Triggers for Origami Scaffolds. 10.2.2.1 Triggering Origami with Temperature

10.2.2.2 Triggering Origami with Electric Fields

10.2.2.3 Triggering Origami with Magnetic Fields

10.2.2.4 Triggering Origami with Light

10.3 DNA Origami Scaffolds for Defined Mechanical Operations

10.3.1 Origami Scaffolds that Dictate the Motility of Elements

10.3.2 Engineering Mechanical Functions of Origami Tiles

10.4 Switchable Interconnected 2D Origami Assemblies

10.5 Dynamic Triggered Switching of Origami for Controlled Release

10.6 Switchable Plasmonic Phenomena with DNA Origami Scaffolds

10.7 Origami‐Guided Organization of Nanoparticles and Proteins

10.8 Conclusions and Perspectives

References

11 The Effect of DNA Boundaries on Enzymatic Reactions

11.1 Introduction

11.2 DNA‐Scaffolded Single Enzymes

11.3 DNA‐Scaffolded Enzyme Cascades

11.4 On the Proximity Model and Other Hypotheses

11.5 Conclusions

Acknowledgments

References

12 The Methods to Assemble Functional Proteins on DNA Scaffold and their Applications

12.1 Introduction

12.2 Overview of the Methods for Arranging Proteins on DNA Scaffolds

12.2.1 Reversible Conjugation between Protein and DNA

12.2.1.1 Biotin‐Avidin

12.2.1.2 Antibody‐Antigen

12.2.1.3 Ni‐NTA‐Hexahistidine

12.2.1.4 Aptamers

12.2.1.5 Apo‐Protein Reconstitution by the Prosthetic Group

12.2.2 Irreversible Conjugation between Protein and DNA

12.2.2.1 Chemical Crosslinking of Protein and DNA via Cross‐Linker

12.2.2.2 Crosslinking of Genetically Fused Protein with Chemically Modified DNA

12.2.2.3 Covalent Conjugation of Genetically Modified Proteins to Unmodified DNA

12.2.2.4 Applications of the Enzyme Assembled DNA Scaffolds

12.3 DNA‐Binding Adaptor for Assembling Proteins on DNA Scaffold and its Application

12.3.1 DNA‐Binding Adaptor for Reversible Assembly of Proteins via Noncovalent Interactions

12.3.2 Modular Adaptors for Covalent Conjugation of Genetically Modified Proteins to Chemically Modified DNA

12.3.3 Application of DNA‐Binding Adaptors for Assembling Proteins on DNA Scaffolds

12.3.3.1 Assembling Protein of Interest on DNA Scaffold in Cell

12.3.3.2 Enzymatic Reaction System on a DNA Scaffold

12.4 Summary

References

13 DNA Origami for Synthetic Biology: An Integrated Gene Logic‐Chip

13.1 Introduction

13.2 Biomolecule Integration on DNA Nanostructure. 13.2.1 Nature Uses “Reaction Field” to Overcome the Cross‐Talk Problem

13.2.2 Synthetic Biology Approach

13.2.3 DNA–Protein Complex

13.2.4 Enzymatic Reaction on DNA Origami for Low‐Molecular‐Weight Substrate

13.3 Gene Expression Control Using DNA Nanostructure. 13.3.1 Enzymatic Reaction on DNA Origami for High‐Molecular‐Weight Substrate

13.3.2 Resolving Synthetic Biology Limitation by DNA Origami‐Based Nano‐Chip

13.3.3 Unique Characters of the Nano‐Chip

13.3.4 Limitation of the Nano‐Chip

13.4 Summary and Perspective

Acknowledgments

References

14 DNA Origami for Molecular Robotics

14.1 DNA Origami as a Stage for DNA Walkers and Robotic Arms

14.2 Nanomechanical DNA Origami

14.3 DNA Origami Used in Combination with Molecular Motors

14.4 Future Perspective

References

15 DNA origami Nanotechnology for the Visualization, Analysis, and Control of Molecular Events with Nanoscale Precision

15.1 Introduction

15.2 Designing of DNA Origami Frames for the Direct Observation of DNA Conformational Changes

15.3 Direct Observation of DNA Structural Changes in the DNA Origami Frame

15.3.1 G‐Quadruplex Formation and Disruption

15.3.2 G‐Quadruplex Formation by the Assembly of Four DNA Strands

15.3.3 Light‐Induced Hybridization and Dehybridization of the Photoswitchable DNA Strands

15.3.4 Direct Observation of B–Z Transition in the Equilibrium State

15.3.5 Topological Control of G‐Quadruplex and I‐Motif Formation in the dsDNA

15.4 Direct Observation and Regulation of Enzyme Reactions in the DNA Origami Frame

15.4.1 Direct Observation and Regulation of Cre‐Mediated DNA Recombination in the DNA Origami Frame

15.4.2 Holiday‐Junction Resolution Mediated by DNA Resolvase

15.4.3 DNA Oxidation in the DNA Demethylation Process Mediated by TET Enzyme

15.4.4 Searching and Recognition of Target Sites by using Photoresponsive Transcription Factor GAL4

15.5 Direct Observation of a Mobile DNA Nanomachine using DNA Origami

15.5.1 A DNA Linear Motor System Created on a DNA Origami System

15.5.2 Single‐Molecule Operation of DNA Motor by using Programmed Instructions

15.5.3 Photo‐Controlled DNA Motor System Constructed on DNA Origami

15.5.4 Photo‐Controlled DNA Rotator System Constructed on DNA Origami

15.6 Limitations of AFM Imaging and Comparison with other Imaging Techniques

15.7 Conclusions and Perspectives

References

16 Stability and Stabilization of DNA Nanostructures in Biomedical Applications

16.1 Threats for DNA Nanostructures

16.1.1 Errors from Nanostructure Synthesis. 16.1.1.1 Missing Strands

16.1.1.2 Oligonucleotide Synthesis Errors

16.1.2 Denaturation of DNA Duplexes. 16.1.2.1 Melting

16.1.2.2 The Role of Cations

16.1.2.3 Influence of pH on Duplex Stability

16.1.3 Backbone Cleavage

16.1.3.1 Acid‐Induced Depurination

16.1.3.2 Base‐Induced Cleavage of RNA

16.1.3.3 Enzymatic Digest

16.1.4 Chemical Damage at the Nucleobases

16.1.4.1 Ultraviolet Radiation

16.1.4.2 Radiative and Oxidative DNA Damage

16.1.4.3 Deamination

16.1.5 DNA Structures for Biological Applications

16.1.5.1 Bioimaging

16.1.5.2 Biosensing

16.1.5.3 Computing

16.1.5.4 Single‐Molecule Biophysics and Mechanobiology

16.1.5.5 Drug Delivery and Gene Therapy

16.1.6 In vitro and in vivo Degradation and Clearance of DNA Structures

16.1.6.1 Common in vitro and in vivo Stability Assays

16.1.6.2 Degradation of DN in in vitro and in vivo

16.1.6.3 Low Mg2+ Conditions

16.1.6.4 Presence of Nucleases

16.1.6.5 Cellular Uptake and Clearance of DNs

16.1.6.6 Immune Response

16.2 Strategies to Protect DNA Origami Structures

16.2.1 Stabilization by Design

16.2.2 Stabilization by Covalent Strategies. 16.2.2.1 Enzymatic Ligation

16.2.2.2 Chemical Crosslinking

16.2.2.3 Photo Crosslinking

16.2.2.4 Base Analogues and Backbone Modification

16.2.3 Stabilization by Non‐Covalent Strategies and Additives

16.2.3.1 Inorganic Materials

16.2.3.2 Proteins

16.2.3.3 Polymer, Peptides, and Polycation Coatings

References

17 DNA Nanostructures for Cancer Diagnosis and Therapy

17.1 Introduction

17.2 DNA Nanostructure‐Based Diagnostics

17.2.1 Nucleic Acid Detection

17.2.2 Protein and Exosome Detection

17.2.3 Tumor Cell Detection

17.2.4 Imaging

17.3 DNA Nanostructure‐Based Drug Delivery

17.3.1 Small Molecules. 17.3.1.1 Doxorubicin

17.3.1.2 Platinum‐Based Drugs

17.3.2 Biologics. 17.3.2.1 CpG

17.3.2.2 RNA

17.3.2.3 Protein

17.3.3 Inorganic Nanoparticles

17.4 Challenges and Prospects

17.4.1 Stability

17.4.1.1 Nucleases

17.4.1.2 Mg2+

17.4.1.3 Shape and Superstructure of DNA Nanostructures

17.4.2 Drug Loading Efficiency

17.4.3 Drug releasing efficiency

17.4.4 Cell Internalization

References

Index. a

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Edited by Masayuki Endo

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Source: Rajendran et al. [60]/with permission of American Chemical Society

(c) Cre‐mediated DNA recombination observed in the DNA frame. Successive HS‐AFM images of the dissociation of the Cre tetramer from the dsDNAs into four Cre monomers and the appearance of a recombinant product. Scanning rate 1.0 frame/s.

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