Description

Book Synopsis
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 moleculesmolecular recognition, self-assembly, programmability, and structural predictabilityhas 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 applicatio

Table of Contents

List of Contributors xiii

Preface xvii

1 DNA Origami Technology: Achievements in the Initial 10 Years 1
Masayuki Endo

1.1 Introduction 1

1.1.1 DNA Nanotechnology Before the Emergence of DNA Origami 3

1.2 Two- Dimensional DNA Origami 3

1.3 Programmed Arrangement of Multiple DNA Origami Components 6

1.4 Three- Dimensional DNA Origami Structures 9

1.5 Modification and Functionalization of 2D DNA Origami Structures 11

1.5.1 Selective Placement of Functional Nanomaterials 11

1.5.2 Selective Placement of Functional Molecules and Proteins via Ligands 13

1.5.3 Distance- Controlled Enzyme Reactions and Photoreactions 13

1.6 Single- Molecule Detection and Sensing using DNA Origami Structures 14

1.6.1 Single- Molecule RNA Detection 14

1.6.2 Single- Molecule Detection of Chemical Reactions 14

1.6.3 Single- Molecule Detection using Mechanical DNA Origami 14

1.6.4 Single- Molecule Sensing using Mechanical DNA Origami 14

1.7 Application to Single Biomolecule AFM Imaging 16

1.7.1 High- Speed AFM- Based Observation of Biomolecules 16

1.7.2 Visualization of DNA Structural Changes in the DNA Nanospace 18

1.7.3 Visualization of the Reaction Events of Enzymes and Proteins in the DNA Nanospace 18

1.8 Single- Molecule Fluorescence Studies 19

1.8.1 Nanoscopic Ruler for Single- Molecule Imaging 19

1.8.2 Kinetics of Binding and Unbinding Events and DNA- PAINT 21

1.8.3 DNA Barcode Imaged by DNA- PAINT 21

1.9 DNA Molecular Machines 22

1.9.1 DNA Assembly Line Constructed on the DNA Origami 22

1.9.2 DNA Spider System Constructed on the DNA Origami 22

1.9.3 DNA Motor System Constructed on the DNA Origami 24

1.10 Selective Incorporation of Nanomaterials and the Applications 24

1.10.1 DNA Origami Plasmonic Structure with Chirality 24

1.10.2 Surface- Enhanced Fluorescence by Gold Nanoparticles and DNA Origami Structure 26

1.10.3 Placement of DNA Origami onto a Fabricated Solid Surface 26

1.11 Dynamic DNA Origami Structures Responsive to External Stimuli 27

1.11.1 DNA Origami Structures Responsive to External Stimuli 27

1.11.2 Stimuli- Responsive DNA Origami Plasmonic Structures 27

1.11.3 Photo- Controlled DNA Origami Plasmonic Structures 27

1.12 Conjugation of DNA Origami to Lipid 29

1.12.1 DNA Origami Channel with Gating 29

1.12.2 DNA Origami Templated Synthesis of Liposomes 29

1.13 DNA Origami for Biological Applications 29

1.13.1 Introduction of DNA Origami into Cells and Functional Expression 29

1.13.2 Drug Release Using the Properties Characteristic for DNA Origami 31

1.13.3 DNA Origami Structures Coated with Lipids and Polymers 32

1.13.4 Nanorobot with Dynamic Mechanism 32

1.13.5 Nanorobot Targeting Tumor In Vivo 32

1.14 Conclusions 33

References 34

2 Wireframe DNA Origami and Its Application as Tools for Molecular Force Generation 41
Marco Lolaico and Björn Högberg

2.1 Introduction 41

2.2 Pre- Origami Wireframe DNA Nanostructures 42

2.3 Hierarchical DNA Origami Wireframe 43

2.4 Entire DNA Origami Design 45

2.5 DNA Origami Wireframe as Tools for Molecular Force Application 50

2.5.1 Introduction 50

2.5.2 Results and Discussion 51

2.6 Conclusions 54

2.6.1 Materials and Methods 54

References 55

3 Capturing Structural Switching and Self- Assembly Events Using High- Speed Atomic Force Microscopy 59
Yuki Suzuki

3.1 Introduction 59

3.2 DNA Origami Nanomachines 60

3.3 Ion- Responsive Mechanical DNA Origami Devices 60

3.4 Photoresponsive Devices 62

3.5 Two- Dimensional Self- Assembly Processes 64

3.6 Sequential Self- Assembly 66

3.7 Photostimulated Assembly and Disassembly 67

3.8 Conclusions and Perspectives 69

References 69

4 Advancement of Computer- Aided Design Software and Simulation Tools for Nucleic Acid Nanostructures and DNA Origami 75
Ibuki Kawamata

4.1 Introduction 75

4.2 General- Purpose Software 76

4.3 Software for Designing Small DNA Nanostructures 78

4.4 Software for Designing DNA Origami 81

4.5 Software for Designing RNA Nanostructures 84

4.6 Software for Designing Base Sequence 84

4.7 Software for Simulating Nucleic Acid Nanostructures 85

4.8 Summary and Future Perspective 86

References 87

5 Dynamic and Mechanical Applications of DNA Nanostructures in Biophysics 101
Melika Shahhosseini, Anjelica Kucinic, Peter Beshay, Wolfgang Pfeifer, and Carlos Castro

5.1 Introduction 101

5.1.1 What Makes DNA a Good Material for Dynamic Applications 101

5.1.2 Rupture Forces 103

5.2 Applications 105

5.2.1 Force Spectroscopy 105

5.2.1.1 Utilizing the Stiffness of DNA for Force Spectroscopy 105

5.2.1.2 Applications that Utilize Rupture Forces 107

5.2.2 DNA Devices that Probe and Control DNA–DNA Interactions 108

5.2.2.1 Detection 108

5.2.2.2 Modulation 111

5.2.3 DNA Devices that Respond to Biomolecules 111

5.2.4 DNA Devices to Study Biological Molecular Motors 116

5.2.5 DNA Walkers 116

5.2.6 DNA Computing 119

5.3 Tools for Quantifying DNA Devices and their Functions 120

5.4 Modeling and Analysis 123

5.5 Conclusion 124

References 124

6 Plasmonic Nanostructures Assembled by DNA Origami 135
Sergio Kogikoski, Jr, Anushree Dutta, and Ilko Bald

6.1 Introduction 135

6.2 Optical Properties of the DNA Origami- Based Plasmonic Nanostructures 135

6.3 Nanoparticle Functionalization with DNA 138

6.4 DNA Origami- Based Plasmonic Assemblies 140

6.5 Surface- Enhanced Raman Scattering (SERS) and Other Plasmonic Effects 143

6.6 Conclusion 152

Acknowledgments 152

References 152

7 Assembly of Nanoparticle Superlattices Using DNA Origami as a Template 155
Sofia Julin, Petteri Piskunen, Mauri A. Kostiainen, and Veikko Linko

7.1 Introduction 155

7.2 Gold Nanoparticles 156

7.2.1 Oligonucleotide- Modified AuNPs 156

7.2.2 Cationic AuNPs 158

7.3 Formation of DNA Origami- Assisted Superlattices 158

7.3.1 Superlattices Formed by Oligonucleotide- Functionalized AuNPs 159

7.3.2 Superlattice Formed by Cationic AuNPs 160

7.4 Characterization of Assemblies 160

7.4.1 Electron Microscopy 161

7.4.2 Small- Angle X- ray Scattering 161

7.5 Conclusions and Future Perspectives 162

Acknowledgments 164

References 164

8 Mechanics of DNA Origami Nanoassemblies 167
Deepak Karna, Jiahao Ji, and Hanbin Mao

8.1 Introduction 167

8.2 Analytical Tools to Investigate Mechanical Properties of Nanoassemblies 168

8.2.1 Optical Tweezers 168

8.2.2 Magnetic Tweezers 169

8.2.3 Atomic Force Microscopy (AFM) 169

8.3 Mechanical Strength of DNA Origami Structures 171

8.4 Applications of Origami Nanostructures by Exploiting their Mechanical Strength 173

8.5 Mechanochemical Properties of DNA Origami 175

8.6 Conclusions 177

References 177

9 3D DNA Origami as Single- Molecule Biophysical Tools for Dissecting Molecular Motor Functions 181
Mitsuhiro Iwaki

9.1 Introduction 181

9.2 DNA Origami Nanospring 181

9.2.1 Design of DNA Origami Nanospring 181

9.2.2 Nanospring Mechanical Properties 182

9.2.3 Application to a Myosin VI Processive Motor 183

9.3 DNA Origami Thick Filament Mimicking Muscle Structure 187

9.3.1 Mystery of Muscle Contraction 187

9.3.2 Design of a DNA Origami- Based Thick Filament 188

9.3.3 High- speed AFM Observation of Force Generation by Myosin 189

9.3.4 High- Speed Darkfield Imaging of Force Generation by Myosin 189

9.4 Perspective 193

References 193

10 Switchable DNA Origami Nanostructures and Their Applications 197
Jianbang Wang, Michael P. O’Hagan, Verena Wulf, and Itamar Willner

10.1 Introduction 197

10.2 Switchable Machines Constructed from DNA Origami Scaffolds 198

10.2.1 Chemical Triggers for Origami Scaffolds 198

10.2.1.1 Triggering Origami Devices with Strand Displacement Reactions 198

10.2.1.2 Triggering Origami with Ion Concentration 200

10.2.1.3 Triggering Origami with Molecular Species 202

10.2.2 Physical Triggers for Origami Scaffolds 204

10.2.2.1 Triggering Origami with Temperature 204

10.2.2.2 Triggering Origami with Electric Fields 206

10.2.2.3 Triggering Origami with Magnetic Fields 206

10.2.2.4 Triggering Origami with Light 208

10.3 DNA Origami Scaffolds for Defined Mechanical Operations 210

10.3.1 Origami Scaffolds that Dictate the Motility of Elements 212

10.3.2 Engineering Mechanical Functions of Origami Tiles 218

10.4 Switchable Interconnected 2D Origami Assemblies 218

10.5 Dynamic Triggered Switching of Origami for Controlled Release 223

10.6 Switchable Plasmonic Phenomena with DNA Origami Scaffolds 227

10.7 Origami- Guided Organization of Nanoparticles and Proteins 234

10.8 Conclusions and Perspectives 238

References 239

11 The Effect of DNA Boundaries on Enzymatic Reactions 241
Richard Kosinski and Barbara Saccà

11.1 Introduction 241

11.2 DNA- Scaffolded Single Enzymes 242

11.3 DNA- Scaffolded Enzyme Cascades 247

11.4 On the Proximity Model and Other Hypotheses 250

11.5 Conclusions 254

Acknowledgments 256

References 256

12 The Methods to Assemble Functional Proteins on DNA Scaffold and their Applications 261
Eiji Nakata, Shiwei Zhang, Huyen Dinh, Peng Lin, and Takashi Morii

12.1 Introduction 261

12.2 Overview of the Methods for Arranging Proteins on DNA Scaffolds 262

12.2.1 Reversible Conjugation between Protein and DNA 263

12.2.1.1 Biotin- Avidin 264

12.2.1.2 Antibody- Antigen 264

12.2.1.3 Ni- NTA- Hexahistidine 266

12.2.1.4 Aptamers 266

12.2.1.5 Apo- Protein Reconstitution by the Prosthetic Group 266

12.2.2 Irreversible Conjugation between Protein and DNA 266

12.2.2.1 Chemical Crosslinking of Protein and DNA via Cross- Linker 267

12.2.2.2 Crosslinking of Genetically Fused Protein with Chemically Modified DNA 267

12.2.2.3 Covalent Conjugation of Genetically Modified Proteins to Unmodified DNA 269

12.2.2.4 Applications of the Enzyme Assembled DNA Scaffolds 269

12.3 DNA- Binding Adaptor for Assembling Proteins on DNA Scaffold and its Application 270

12.3.1 DNA- Binding Adaptor for Reversible Assembly of Proteins via Noncovalent Interactions 270

12.3.2 Modular Adaptors for Covalent Conjugation of Genetically Modified Proteins to Chemically Modified DNA 272

12.3.3 Application of DNA- Binding Adaptors for Assembling Proteins on DNA Scaffolds 275

12.3.3.1 Assembling Protein of Interest on DNA Scaffold in Cell 275

12.3.3.2 Enzymatic Reaction System on a DNA Scaffold 275

12.4 Summary 278

References 278

13 DNA Origami for Synthetic Biology: An Integrated Gene Logic- Chip 281
Hisashi Tadakuma

13.1 Introduction 281

13.2 Biomolecule Integration on DNA Nanostructure 281

13.2.1 Nature Uses “Reaction Field” to Overcome the Cross- Talk Problem 281

13.2.2 Synthetic Biology Approach 282

13.2.3 DNA–Protein Complex 282

13.2.4 Enzymatic Reaction on DNA Origami for Low- Molecular- Weight Substrate 284

13.3 Gene Expression Control Using DNA Nanostructure 285

13.3.1 Enzymatic Reaction on DNA Origami for High- Molecular- Weight Substrate 285

13.3.2 Resolving Synthetic Biology Limitation by DNA Origami- Based Nano- Chip 286

13.3.3 Unique Characters of the Nano- Chip 288

13.3.4 Limitation of the Nano- Chip 292

13.4 Summary and Perspective 292

Acknowledgments 293

References 293

14 DNA Origami for Molecular Robotics 297
Akinori Kuzuya

14.1 DNA Origami as a Stage for DNA Walkers and Robotic Arms 297

14.2 Nanomechanical DNA Origami 298

14.3 DNA Origami Used in Combination with Molecular Motors 300

14.4 Future Perspective 301

References 303

15 DNA origami Nanotechnology for the Visualization, Analysis, and Control of Molecular Events with Nanoscale Precision 305
Xiwen Xing and Masayuki Endo

15.1 Introduction 305

15.2 Designing of DNA Origami Frames for the Direct Observation of DNA Conformational Changes 308

15.3 Direct Observation of DNA Structural Changes in the DNA Origami Frame 308

15.3.1 G- Quadruplex Formation and Disruption 308

15.3.2 G- Quadruplex Formation by the Assembly of Four DNA Strands 309

15.3.3 Light- Induced Hybridization and Dehybridization of the Photoswitchable DNA Strands 309

15.3.4 Direct Observation of B–Z Transition in the Equilibrium State 312

15.3.5 Topological Control of G- Quadruplex and I- Motif Formation in the dsDNA 314

15.4 Direct Observation and Regulation of Enzyme Reactions in the DNA Origami Frame 315

15.4.1 Direct Observation and Regulation of Cre- Mediated DNA Recombination in the DNA Origami Frame 315

15.4.2 Holiday- Junction Resolution Mediated by DNA Resolvase 317

15.4.3 DNA Oxidation in the DNA Demethylation Process Mediated by TET Enzyme 317

15.4.4 Searching and Recognition of Target Sites by using Photoresponsive Transcription Factor GAL 4 319

15.5 Direct Observation of a Mobile DNA Nanomachine using DNA Origami 321

15.5.1 A DNA Linear Motor System Created on a DNA Origami System 321

15.5.2 Single- Molecule Operation of DNA Motor by using Programmed Instructions 321

15.5.3 Photo- Controlled DNA Motor System Constructed on DNA Origami 324

15.5.4 Photo- Controlled DNA Rotator System Constructed on DNA Origami 324

15.6 Limitations of AFM Imaging and Comparison with other Imaging Techniques 326

15.7 Conclusions and Perspectives 326

References 327

16 Stability and Stabilization of DNA Nanostructures in Biomedical Applications 333
Soumya Chandrasekhar, Praneetha Sundar Prakash, and Thorsten- Lars Schmidt

16.1 Threats for DNA Nanostructures 333

16.1.1 Errors from Nanostructure Synthesis 334

16.1.1.1 Missing Strands 334

16.1.1.2 Oligonucleotide Synthesis Errors 335

16.1.2 Denaturation of DNA Duplexes 336

16.1.2.1 Melting 336

16.1.2.2 The Role of Cations 336

16.1.2.3 Influence of pH on Duplex Stability 337

16.1.3 Backbone Cleavage 337

16.1.3.1 Acid- Induced Depurination 337

16.1.3.2 Base- Induced Cleavage of RNA 338

16.1.3.3 Enzymatic Digest 338

16.1.4 Chemical Damage at the Nucleobases 339

16.1.4.1 Ultraviolet Radiation 339

16.1.4.2 Radiative and Oxidative DNA Damage 340

16.1.4.3 Deamination 340

16.1.5 DNA Structures for Biological Applications 341

16.1.5.1 Bioimaging 341

16.1.5.2 Biosensing 341

16.1.5.3 Computing 341

16.1.5.4 Single- Molecule Biophysics and Mechanobiology 343

16.1.5.5 Drug Delivery and Gene Therapy 343

16.1.6 In vitro and In vivo Degradation and Clearance of DNA Structures 343

16.1.6.1 Common in vitro and in vivo Stability Assays 344

16.1.6.2 Degradation of DN in in vitro and in vivo 344

16.1.6.3 Low Mg2+ Conditions 346

16.1.6.4 Presence of Nucleases 346

16.1.6.5 Cellular Uptake and Clearance of DNs 347

16.1.6.6 Immune Response 348

16.2 Strategies to Protect DNA Origami Structures 349

16.2.1 Stabilization by Design 349

16.2.2 Stabilization by Covalent Strategies 351

16.2.2.1 Enzymatic Ligation 351

16.2.2.2 Chemical Crosslinking 352

16.2.2.3 Photo Crosslinking 354

16.2.2.4 Base Analogues and Backbone Modification 356

16.2.3 Stabilization by Non- Covalent Strategies and Additives 356

16.2.3.1 Inorganic Materials 356

16.2.3.2 Proteins 358

16.2.3.3 Polymer, Peptides, and Polycation Coatings 358

References 362

17 DNA Nanostructures for Cancer Diagnosis and Therapy 379
Zhe Li and Yonggang Ke

17.1 Introduction 379

17.2 DNA Nanostructure- Based Diagnostics 380

17.2.1 Nucleic Acid Detection 380

17.2.2 Protein and Exosome Detection 382

17.2.3 Tumor Cell Detection 384

17.2.4 Imaging 385

17.3 DNA Nanostructure- Based Drug Delivery 386

17.3.1 Small Molecules 386

17.3.1.1 Doxorubicin 386

17.3.1.2 Platinum- Based Drugs 387

17.3.2 Biologics 389

17.3.2.1 CpG 389

17.3.2.2 RNA 390

17.3.2.3 Protein 392

17.3.3 Inorganic Nanoparticles 393

17.4 Challenges and Prospects 394

17.4.1 Stability 394

17.4.1.1 Nucleases 395

17.4.1.2 Mg2+ 395

17.4.1.3 Shape and Superstructure of DNA Nanostructures 396

17.4.2 Drug Loading Efficiency 396

17.4.3 Drug releasing efficiency 397

17.4.4 Cell Internalization 398

References 400

Index 411

DNA Origami

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    Publisher: John Wiley & Sons Inc
    Publication Date: 10/05/2022
    ISBN13: 9781119682547, 978-1119682547
    ISBN10: 1119682541

    Description

    Book Synopsis
    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 moleculesmolecular recognition, self-assembly, programmability, and structural predictabilityhas 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 applicatio

    Table of Contents

    List of Contributors xiii

    Preface xvii

    1 DNA Origami Technology: Achievements in the Initial 10 Years 1
    Masayuki Endo

    1.1 Introduction 1

    1.1.1 DNA Nanotechnology Before the Emergence of DNA Origami 3

    1.2 Two- Dimensional DNA Origami 3

    1.3 Programmed Arrangement of Multiple DNA Origami Components 6

    1.4 Three- Dimensional DNA Origami Structures 9

    1.5 Modification and Functionalization of 2D DNA Origami Structures 11

    1.5.1 Selective Placement of Functional Nanomaterials 11

    1.5.2 Selective Placement of Functional Molecules and Proteins via Ligands 13

    1.5.3 Distance- Controlled Enzyme Reactions and Photoreactions 13

    1.6 Single- Molecule Detection and Sensing using DNA Origami Structures 14

    1.6.1 Single- Molecule RNA Detection 14

    1.6.2 Single- Molecule Detection of Chemical Reactions 14

    1.6.3 Single- Molecule Detection using Mechanical DNA Origami 14

    1.6.4 Single- Molecule Sensing using Mechanical DNA Origami 14

    1.7 Application to Single Biomolecule AFM Imaging 16

    1.7.1 High- Speed AFM- Based Observation of Biomolecules 16

    1.7.2 Visualization of DNA Structural Changes in the DNA Nanospace 18

    1.7.3 Visualization of the Reaction Events of Enzymes and Proteins in the DNA Nanospace 18

    1.8 Single- Molecule Fluorescence Studies 19

    1.8.1 Nanoscopic Ruler for Single- Molecule Imaging 19

    1.8.2 Kinetics of Binding and Unbinding Events and DNA- PAINT 21

    1.8.3 DNA Barcode Imaged by DNA- PAINT 21

    1.9 DNA Molecular Machines 22

    1.9.1 DNA Assembly Line Constructed on the DNA Origami 22

    1.9.2 DNA Spider System Constructed on the DNA Origami 22

    1.9.3 DNA Motor System Constructed on the DNA Origami 24

    1.10 Selective Incorporation of Nanomaterials and the Applications 24

    1.10.1 DNA Origami Plasmonic Structure with Chirality 24

    1.10.2 Surface- Enhanced Fluorescence by Gold Nanoparticles and DNA Origami Structure 26

    1.10.3 Placement of DNA Origami onto a Fabricated Solid Surface 26

    1.11 Dynamic DNA Origami Structures Responsive to External Stimuli 27

    1.11.1 DNA Origami Structures Responsive to External Stimuli 27

    1.11.2 Stimuli- Responsive DNA Origami Plasmonic Structures 27

    1.11.3 Photo- Controlled DNA Origami Plasmonic Structures 27

    1.12 Conjugation of DNA Origami to Lipid 29

    1.12.1 DNA Origami Channel with Gating 29

    1.12.2 DNA Origami Templated Synthesis of Liposomes 29

    1.13 DNA Origami for Biological Applications 29

    1.13.1 Introduction of DNA Origami into Cells and Functional Expression 29

    1.13.2 Drug Release Using the Properties Characteristic for DNA Origami 31

    1.13.3 DNA Origami Structures Coated with Lipids and Polymers 32

    1.13.4 Nanorobot with Dynamic Mechanism 32

    1.13.5 Nanorobot Targeting Tumor In Vivo 32

    1.14 Conclusions 33

    References 34

    2 Wireframe DNA Origami and Its Application as Tools for Molecular Force Generation 41
    Marco Lolaico and Björn Högberg

    2.1 Introduction 41

    2.2 Pre- Origami Wireframe DNA Nanostructures 42

    2.3 Hierarchical DNA Origami Wireframe 43

    2.4 Entire DNA Origami Design 45

    2.5 DNA Origami Wireframe as Tools for Molecular Force Application 50

    2.5.1 Introduction 50

    2.5.2 Results and Discussion 51

    2.6 Conclusions 54

    2.6.1 Materials and Methods 54

    References 55

    3 Capturing Structural Switching and Self- Assembly Events Using High- Speed Atomic Force Microscopy 59
    Yuki Suzuki

    3.1 Introduction 59

    3.2 DNA Origami Nanomachines 60

    3.3 Ion- Responsive Mechanical DNA Origami Devices 60

    3.4 Photoresponsive Devices 62

    3.5 Two- Dimensional Self- Assembly Processes 64

    3.6 Sequential Self- Assembly 66

    3.7 Photostimulated Assembly and Disassembly 67

    3.8 Conclusions and Perspectives 69

    References 69

    4 Advancement of Computer- Aided Design Software and Simulation Tools for Nucleic Acid Nanostructures and DNA Origami 75
    Ibuki Kawamata

    4.1 Introduction 75

    4.2 General- Purpose Software 76

    4.3 Software for Designing Small DNA Nanostructures 78

    4.4 Software for Designing DNA Origami 81

    4.5 Software for Designing RNA Nanostructures 84

    4.6 Software for Designing Base Sequence 84

    4.7 Software for Simulating Nucleic Acid Nanostructures 85

    4.8 Summary and Future Perspective 86

    References 87

    5 Dynamic and Mechanical Applications of DNA Nanostructures in Biophysics 101
    Melika Shahhosseini, Anjelica Kucinic, Peter Beshay, Wolfgang Pfeifer, and Carlos Castro

    5.1 Introduction 101

    5.1.1 What Makes DNA a Good Material for Dynamic Applications 101

    5.1.2 Rupture Forces 103

    5.2 Applications 105

    5.2.1 Force Spectroscopy 105

    5.2.1.1 Utilizing the Stiffness of DNA for Force Spectroscopy 105

    5.2.1.2 Applications that Utilize Rupture Forces 107

    5.2.2 DNA Devices that Probe and Control DNA–DNA Interactions 108

    5.2.2.1 Detection 108

    5.2.2.2 Modulation 111

    5.2.3 DNA Devices that Respond to Biomolecules 111

    5.2.4 DNA Devices to Study Biological Molecular Motors 116

    5.2.5 DNA Walkers 116

    5.2.6 DNA Computing 119

    5.3 Tools for Quantifying DNA Devices and their Functions 120

    5.4 Modeling and Analysis 123

    5.5 Conclusion 124

    References 124

    6 Plasmonic Nanostructures Assembled by DNA Origami 135
    Sergio Kogikoski, Jr, Anushree Dutta, and Ilko Bald

    6.1 Introduction 135

    6.2 Optical Properties of the DNA Origami- Based Plasmonic Nanostructures 135

    6.3 Nanoparticle Functionalization with DNA 138

    6.4 DNA Origami- Based Plasmonic Assemblies 140

    6.5 Surface- Enhanced Raman Scattering (SERS) and Other Plasmonic Effects 143

    6.6 Conclusion 152

    Acknowledgments 152

    References 152

    7 Assembly of Nanoparticle Superlattices Using DNA Origami as a Template 155
    Sofia Julin, Petteri Piskunen, Mauri A. Kostiainen, and Veikko Linko

    7.1 Introduction 155

    7.2 Gold Nanoparticles 156

    7.2.1 Oligonucleotide- Modified AuNPs 156

    7.2.2 Cationic AuNPs 158

    7.3 Formation of DNA Origami- Assisted Superlattices 158

    7.3.1 Superlattices Formed by Oligonucleotide- Functionalized AuNPs 159

    7.3.2 Superlattice Formed by Cationic AuNPs 160

    7.4 Characterization of Assemblies 160

    7.4.1 Electron Microscopy 161

    7.4.2 Small- Angle X- ray Scattering 161

    7.5 Conclusions and Future Perspectives 162

    Acknowledgments 164

    References 164

    8 Mechanics of DNA Origami Nanoassemblies 167
    Deepak Karna, Jiahao Ji, and Hanbin Mao

    8.1 Introduction 167

    8.2 Analytical Tools to Investigate Mechanical Properties of Nanoassemblies 168

    8.2.1 Optical Tweezers 168

    8.2.2 Magnetic Tweezers 169

    8.2.3 Atomic Force Microscopy (AFM) 169

    8.3 Mechanical Strength of DNA Origami Structures 171

    8.4 Applications of Origami Nanostructures by Exploiting their Mechanical Strength 173

    8.5 Mechanochemical Properties of DNA Origami 175

    8.6 Conclusions 177

    References 177

    9 3D DNA Origami as Single- Molecule Biophysical Tools for Dissecting Molecular Motor Functions 181
    Mitsuhiro Iwaki

    9.1 Introduction 181

    9.2 DNA Origami Nanospring 181

    9.2.1 Design of DNA Origami Nanospring 181

    9.2.2 Nanospring Mechanical Properties 182

    9.2.3 Application to a Myosin VI Processive Motor 183

    9.3 DNA Origami Thick Filament Mimicking Muscle Structure 187

    9.3.1 Mystery of Muscle Contraction 187

    9.3.2 Design of a DNA Origami- Based Thick Filament 188

    9.3.3 High- speed AFM Observation of Force Generation by Myosin 189

    9.3.4 High- Speed Darkfield Imaging of Force Generation by Myosin 189

    9.4 Perspective 193

    References 193

    10 Switchable DNA Origami Nanostructures and Their Applications 197
    Jianbang Wang, Michael P. O’Hagan, Verena Wulf, and Itamar Willner

    10.1 Introduction 197

    10.2 Switchable Machines Constructed from DNA Origami Scaffolds 198

    10.2.1 Chemical Triggers for Origami Scaffolds 198

    10.2.1.1 Triggering Origami Devices with Strand Displacement Reactions 198

    10.2.1.2 Triggering Origami with Ion Concentration 200

    10.2.1.3 Triggering Origami with Molecular Species 202

    10.2.2 Physical Triggers for Origami Scaffolds 204

    10.2.2.1 Triggering Origami with Temperature 204

    10.2.2.2 Triggering Origami with Electric Fields 206

    10.2.2.3 Triggering Origami with Magnetic Fields 206

    10.2.2.4 Triggering Origami with Light 208

    10.3 DNA Origami Scaffolds for Defined Mechanical Operations 210

    10.3.1 Origami Scaffolds that Dictate the Motility of Elements 212

    10.3.2 Engineering Mechanical Functions of Origami Tiles 218

    10.4 Switchable Interconnected 2D Origami Assemblies 218

    10.5 Dynamic Triggered Switching of Origami for Controlled Release 223

    10.6 Switchable Plasmonic Phenomena with DNA Origami Scaffolds 227

    10.7 Origami- Guided Organization of Nanoparticles and Proteins 234

    10.8 Conclusions and Perspectives 238

    References 239

    11 The Effect of DNA Boundaries on Enzymatic Reactions 241
    Richard Kosinski and Barbara Saccà

    11.1 Introduction 241

    11.2 DNA- Scaffolded Single Enzymes 242

    11.3 DNA- Scaffolded Enzyme Cascades 247

    11.4 On the Proximity Model and Other Hypotheses 250

    11.5 Conclusions 254

    Acknowledgments 256

    References 256

    12 The Methods to Assemble Functional Proteins on DNA Scaffold and their Applications 261
    Eiji Nakata, Shiwei Zhang, Huyen Dinh, Peng Lin, and Takashi Morii

    12.1 Introduction 261

    12.2 Overview of the Methods for Arranging Proteins on DNA Scaffolds 262

    12.2.1 Reversible Conjugation between Protein and DNA 263

    12.2.1.1 Biotin- Avidin 264

    12.2.1.2 Antibody- Antigen 264

    12.2.1.3 Ni- NTA- Hexahistidine 266

    12.2.1.4 Aptamers 266

    12.2.1.5 Apo- Protein Reconstitution by the Prosthetic Group 266

    12.2.2 Irreversible Conjugation between Protein and DNA 266

    12.2.2.1 Chemical Crosslinking of Protein and DNA via Cross- Linker 267

    12.2.2.2 Crosslinking of Genetically Fused Protein with Chemically Modified DNA 267

    12.2.2.3 Covalent Conjugation of Genetically Modified Proteins to Unmodified DNA 269

    12.2.2.4 Applications of the Enzyme Assembled DNA Scaffolds 269

    12.3 DNA- Binding Adaptor for Assembling Proteins on DNA Scaffold and its Application 270

    12.3.1 DNA- Binding Adaptor for Reversible Assembly of Proteins via Noncovalent Interactions 270

    12.3.2 Modular Adaptors for Covalent Conjugation of Genetically Modified Proteins to Chemically Modified DNA 272

    12.3.3 Application of DNA- Binding Adaptors for Assembling Proteins on DNA Scaffolds 275

    12.3.3.1 Assembling Protein of Interest on DNA Scaffold in Cell 275

    12.3.3.2 Enzymatic Reaction System on a DNA Scaffold 275

    12.4 Summary 278

    References 278

    13 DNA Origami for Synthetic Biology: An Integrated Gene Logic- Chip 281
    Hisashi Tadakuma

    13.1 Introduction 281

    13.2 Biomolecule Integration on DNA Nanostructure 281

    13.2.1 Nature Uses “Reaction Field” to Overcome the Cross- Talk Problem 281

    13.2.2 Synthetic Biology Approach 282

    13.2.3 DNA–Protein Complex 282

    13.2.4 Enzymatic Reaction on DNA Origami for Low- Molecular- Weight Substrate 284

    13.3 Gene Expression Control Using DNA Nanostructure 285

    13.3.1 Enzymatic Reaction on DNA Origami for High- Molecular- Weight Substrate 285

    13.3.2 Resolving Synthetic Biology Limitation by DNA Origami- Based Nano- Chip 286

    13.3.3 Unique Characters of the Nano- Chip 288

    13.3.4 Limitation of the Nano- Chip 292

    13.4 Summary and Perspective 292

    Acknowledgments 293

    References 293

    14 DNA Origami for Molecular Robotics 297
    Akinori Kuzuya

    14.1 DNA Origami as a Stage for DNA Walkers and Robotic Arms 297

    14.2 Nanomechanical DNA Origami 298

    14.3 DNA Origami Used in Combination with Molecular Motors 300

    14.4 Future Perspective 301

    References 303

    15 DNA origami Nanotechnology for the Visualization, Analysis, and Control of Molecular Events with Nanoscale Precision 305
    Xiwen Xing and Masayuki Endo

    15.1 Introduction 305

    15.2 Designing of DNA Origami Frames for the Direct Observation of DNA Conformational Changes 308

    15.3 Direct Observation of DNA Structural Changes in the DNA Origami Frame 308

    15.3.1 G- Quadruplex Formation and Disruption 308

    15.3.2 G- Quadruplex Formation by the Assembly of Four DNA Strands 309

    15.3.3 Light- Induced Hybridization and Dehybridization of the Photoswitchable DNA Strands 309

    15.3.4 Direct Observation of B–Z Transition in the Equilibrium State 312

    15.3.5 Topological Control of G- Quadruplex and I- Motif Formation in the dsDNA 314

    15.4 Direct Observation and Regulation of Enzyme Reactions in the DNA Origami Frame 315

    15.4.1 Direct Observation and Regulation of Cre- Mediated DNA Recombination in the DNA Origami Frame 315

    15.4.2 Holiday- Junction Resolution Mediated by DNA Resolvase 317

    15.4.3 DNA Oxidation in the DNA Demethylation Process Mediated by TET Enzyme 317

    15.4.4 Searching and Recognition of Target Sites by using Photoresponsive Transcription Factor GAL 4 319

    15.5 Direct Observation of a Mobile DNA Nanomachine using DNA Origami 321

    15.5.1 A DNA Linear Motor System Created on a DNA Origami System 321

    15.5.2 Single- Molecule Operation of DNA Motor by using Programmed Instructions 321

    15.5.3 Photo- Controlled DNA Motor System Constructed on DNA Origami 324

    15.5.4 Photo- Controlled DNA Rotator System Constructed on DNA Origami 324

    15.6 Limitations of AFM Imaging and Comparison with other Imaging Techniques 326

    15.7 Conclusions and Perspectives 326

    References 327

    16 Stability and Stabilization of DNA Nanostructures in Biomedical Applications 333
    Soumya Chandrasekhar, Praneetha Sundar Prakash, and Thorsten- Lars Schmidt

    16.1 Threats for DNA Nanostructures 333

    16.1.1 Errors from Nanostructure Synthesis 334

    16.1.1.1 Missing Strands 334

    16.1.1.2 Oligonucleotide Synthesis Errors 335

    16.1.2 Denaturation of DNA Duplexes 336

    16.1.2.1 Melting 336

    16.1.2.2 The Role of Cations 336

    16.1.2.3 Influence of pH on Duplex Stability 337

    16.1.3 Backbone Cleavage 337

    16.1.3.1 Acid- Induced Depurination 337

    16.1.3.2 Base- Induced Cleavage of RNA 338

    16.1.3.3 Enzymatic Digest 338

    16.1.4 Chemical Damage at the Nucleobases 339

    16.1.4.1 Ultraviolet Radiation 339

    16.1.4.2 Radiative and Oxidative DNA Damage 340

    16.1.4.3 Deamination 340

    16.1.5 DNA Structures for Biological Applications 341

    16.1.5.1 Bioimaging 341

    16.1.5.2 Biosensing 341

    16.1.5.3 Computing 341

    16.1.5.4 Single- Molecule Biophysics and Mechanobiology 343

    16.1.5.5 Drug Delivery and Gene Therapy 343

    16.1.6 In vitro and In vivo Degradation and Clearance of DNA Structures 343

    16.1.6.1 Common in vitro and in vivo Stability Assays 344

    16.1.6.2 Degradation of DN in in vitro and in vivo 344

    16.1.6.3 Low Mg2+ Conditions 346

    16.1.6.4 Presence of Nucleases 346

    16.1.6.5 Cellular Uptake and Clearance of DNs 347

    16.1.6.6 Immune Response 348

    16.2 Strategies to Protect DNA Origami Structures 349

    16.2.1 Stabilization by Design 349

    16.2.2 Stabilization by Covalent Strategies 351

    16.2.2.1 Enzymatic Ligation 351

    16.2.2.2 Chemical Crosslinking 352

    16.2.2.3 Photo Crosslinking 354

    16.2.2.4 Base Analogues and Backbone Modification 356

    16.2.3 Stabilization by Non- Covalent Strategies and Additives 356

    16.2.3.1 Inorganic Materials 356

    16.2.3.2 Proteins 358

    16.2.3.3 Polymer, Peptides, and Polycation Coatings 358

    References 362

    17 DNA Nanostructures for Cancer Diagnosis and Therapy 379
    Zhe Li and Yonggang Ke

    17.1 Introduction 379

    17.2 DNA Nanostructure- Based Diagnostics 380

    17.2.1 Nucleic Acid Detection 380

    17.2.2 Protein and Exosome Detection 382

    17.2.3 Tumor Cell Detection 384

    17.2.4 Imaging 385

    17.3 DNA Nanostructure- Based Drug Delivery 386

    17.3.1 Small Molecules 386

    17.3.1.1 Doxorubicin 386

    17.3.1.2 Platinum- Based Drugs 387

    17.3.2 Biologics 389

    17.3.2.1 CpG 389

    17.3.2.2 RNA 390

    17.3.2.3 Protein 392

    17.3.3 Inorganic Nanoparticles 393

    17.4 Challenges and Prospects 394

    17.4.1 Stability 394

    17.4.1.1 Nucleases 395

    17.4.1.2 Mg2+ 395

    17.4.1.3 Shape and Superstructure of DNA Nanostructures 396

    17.4.2 Drug Loading Efficiency 396

    17.4.3 Drug releasing efficiency 397

    17.4.4 Cell Internalization 398

    References 400

    Index 411

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