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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